Biomarkers for the diagnosis of invasive fungal infections
12181474 · 2024-12-31
Assignee
Inventors
- Janin Schulte (Hennigsdorf, DE)
- Anne Incamps (Saint Victor la Coste, FR)
- Markus Alexander Weigand (Wettenberg, DE)
- Thorsten Brenner (Mörlenbach, DE)
Cpc classification
International classification
Abstract
The present invention relates to a method for the diagnosis, prognosis, risk assessment, risk stratification, monitoring, therapy guidance and/or therapy control of a fungal infection, in particular invasive fungal infections (IFI) and/or the ruling in or ruling out of an fungal infection and/or the differential diagnosis of a fungal colonization vs. an invasive fungal infection in a subject, wherein in particular the subject has an increased risk of getting or having a fungal infection and/or the subject is in a critical disease state, particularly has an existing infection and/or a state of sepsis, particularly a septic shock. The method of the invention comprises determining the level of at least one marker selected from the group of ICAM1, AHSG, CPN1, FABP1, HRG, PIGR, RAP1A, THBS1, VCL, ET-1. Furthermore, the invention relates to a diagnostic assay and a kit for carrying out the method.
Claims
1. A method for treating an acute invasive fungal infection in a subject, comprising the step of measuring a level of intercellular adhesion molecule 1 (ICAM1) in a sample from the subject, and administering an effective amount of an antifungal agent to the subject, wherein the level of ICAM1 is increased 1.5 fold as compared to the level of ICAM1 in a control and indicates the presence of the acute invasive fungal infection in the subject, thereby treating the acute fungal infection in the subject, wherein the subject has immune suppression, an impaired immune response or a dysregulated immune response.
2. The method of claim 1, wherein the method further comprises measuring a level of alpha-2-HS-glycoprotein (AHSG), carboxypeptidase N catalytic chain 1 (CPN1), fatty-acid binding protein 1 (FABP1), histidine rich glycoprotein (HRG), polymeric immunoglobulin receptor (PIGR), ras-related protein 1 (RAP1), thrombospondin-1 (THBS1), vinculin (VCL), endothelin 1 (ET-1), or any combination thereof in a sample from the subject.
3. The method of claim 1, wherein the method further comprises measuring a level of a) THBS1; b) VCL; or c) THBS1 and VCL in a sample from the subject.
4. The method of claim 1, wherein the acute invasive fungal infection is a systemic fungal infection, a fungemia or a multifocal infection.
5. The method of claim 1, wherein the level of ICAM1 is determined after the subject is diagnosed of having a fungal infection, after the subject is diagnosed to be at risk for mortality from a fungal infection, and/or after admission of the subject to a medical site.
6. The method of claim 1 wherein the level of ICAM1 is compared to a reference value, and wherein a level of ICAM1 in the sample, above the reference value, indicates the presence of the acute invasive fungal infection in the subject.
7. The method of claim 6, wherein the reference value is a level of ICAM1 in one or more samples of a reference subject or a population of reference subjects without the acute invasive fungal infection and without a fungal colonization.
8. The method of claim 6, wherein the reference value is a level of ICAM1 in one or more samples of a reference subject or a population of reference subjects without the acute invasive fungal infection and having a fungal colonization.
9. The method of claim 1, wherein the subject is at risk for mortality from a fungal infection.
10. The method of claim 1, wherein the subject is a subject after organ transplantation.
11. The method of claim 9, wherein the subject is (i) a patient having at least one chronic or acute viral or bacterial infection; (ii) a patient having a mixed bacterial and viral infection; or (iii) a patient having an immune suppression, impaired immune response or dysregulated immune system.
12. The method of claim 1, wherein the method further comprises measuring a level of C-reactive protein (CRP), cytokines, procalcitonin (PCT), tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), neutrophil gelatinase-associated lipocalin (NGAL), interferon-induced GTP-binding protein Mx1 (MX1), pancreatic stone protein (PSP), atrial natriuretic peptide, arginine vasopressin, angiotensin II, glucans, interferon gamma, or any combination thereof, in a sample from the subject.
13. The method of claim 12, wherein the method further comprises measuring a level of procalcitonin (PCT) in a sample from the subject.
14. The method of claim 1, wherein the level of ICAM1 is measured by mass spectrometry or in an immunoassay.
15. The method of claim 1, wherein the subject is a liver transplant recipient.
16. The method of claim 1, wherein the method further comprises measuring a level of one or more clinical scores, wherein: the one or more clinical scores comprise sequential organ failure assessment score (SOFA), simplified acute physiology score (SAPSII score), the Acute Physiology and Chronic Health Evaluation II (APACHE II) score, the Pneumonia Severity Index (PSI) score, or any combination thereof.
17. The method of claim 1, wherein the method further comprises measuring a level of one or more clinical parameters, wherein: the one or more clinical parameters comprise age, gender, family history, ethnicity, body weight, body mass index (BMI), systolic blood pressure, diastolic blood pressure, heart rate, temperature, duration of a medicinal intervention, surgical procedures, medication, or any combination thereof.
18. The method of claim 1, wherein the method further comprises measuring a level of one or more infection parameters, wherein: the one or more infection parameters comprise leukocyte count, neutrophil count, or a combination thereof.
19. The method of claim 1, wherein the acute invasive fungal infection is caused by a Candida spp., an Aspergillus spp., or a combination thereof.
20. The method of claim 1, wherein the subject has Systemic Inflammatory Response Syndrome (SIRS) or sepsis.
21. The method of claim 1, wherein the antifungal agent comprises a polyene antifungal drug, an echinocandin, an azole antifungal drug, an allylamine antifungal drug, a morpholine antifungal drug, or an anti-metabolite antifungal drug.
22. A method for treating an acute invasive fungal infection in a subject, comprising the step of measuring a level of intercellular adhesion molecule 1 (ICAM1) in a sample from the subject, and administering an effective amount of an antifungal agent to the subject, wherein the level of ICAM1 is increased 1.5 fold as compared to the level of ICAM1 in a control and indicates the presence of the acute invasive fungal infection in the subject, thereby treating the subject, and wherein: a) the fungal infection is caused by a Candida spp., an Aspergillus spp., or a combination thereof, b) the subject has Systemic Inflammatory Response Syndrome (SIRS) or sepsis; c) the antifungal agent comprises a polyene antifungal drug, an echinocandin, an azole antifungal drug, an allylamine antifungal drug, a morpholine antifungal drug, or an anti-metabolite antifungal drug; and d) the control is the amount of ICAM1 in a sample from a subject with fungal colonization or the amount of ICAM1 is a sample from a subject without a fungal infection.
23. The method of claim 21, wherein the subject has a liver transplant or has septic shock.
Description
DESCRIPTION OF THE DRAWINGS
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(73) The following examples and figures serve for a more detailed explanation of the invention, but without restricting the invention to these examples and figures.
EXAMPLES
Example 1: IFI in the Context of Sepsis, Especially Patients with Septic Shock
(74) Study design: The observational clinical study was approved by the local ethics committee (Ethics Committee of the Medical Faculty of Heidelberg, Trial Code No. S-097/2013/German Clinical Trials Register: DRKS00005463) and was conducted in the surgical intensive care unit of Heidelberg University Hospital, Germany between November 2013 and January 2015. All study patients or their legal designees gave written informed consent. In total 50 patients suffering from septic shock according to the criteria of the Surviving Sepsis Campaign: International Guidelines for Management of Severe Sepsis and Septic Shock 2012 were enrolled in this study (Dellinger et al. Crit Care Med. 2012 41:580-637; Romani. Nat Rev Immunol. 2004 4:1-23); Schroeder M et al. Crit Care. 2016 20:139; Zedek D C et al. J Clin Microbiol. 2006 44:1601).
(75) Blood sample were collected at sepsis onset (T0) and 1 day (T1), 2 days (T2), 7 days (T3) 14 days (T4), 21 days (T5) and 28 days (T6) afterwards. Relevant baseline data (demographic data, primary site of infection), clinical data (disease severity scores, such as Simplified Acute Physiology Score (SAPS II), Sequential Organ Failure Assessment Score (SOFA) and Acute Physiology Health Evaluation score (APACHE II), surgical procedures, antifungal therapy, outcome parameters) as well as routine infection parameters (e.g. leukocytes, C-reactive protein (CRP), procalcitonin (PCT), body temperature) were collected (Table 1).
(76) Immunoassays. Plasma concentrations of -D-glucan (BD) were measured using the Glucatell-Kit (Pyroquant Diagnostik GmbH) according to the manufacturer's instructions. In al patients, concentrations of Galactomannan (GM) were measured using an enzyme-linked immunoassay (Platelia Aspergillus AG, Biorad, and Munich) in plasma samples at all time points. Concentrations of GM in bronchoalveolar lavage fluid (BALF) were measured using the same technique, however only in selected cases of suspected invasive aspergillosis (IA). The following GM concentrations were used as cut-off values: Plasma >0.5, BALF>1.0
(77) The biomarkers ICAM1, AHSG, CPN1, FABP1, HRG, PIGR, RAP1, THBS1, VCL were measured in quantitative selected reaction monitoring (SRM) assays by LC-MS/MS technology (TSQ Quantiva mass spectrometer (MS); ThermoFisher Scientific). PCT and ET-1 were measured by the automated immunoassay platform Kryptor Brahms PCT.
SEQUENCE LISTING
(78) The Sequence Listing is submitted as an ASCII text file [10278-105441-01_Sequence_Listing.txt, Nov. 23, 2020, 73.6 KB], which is incorporated by reference herein.
(79) Clinical Microbiology.
(80) Blood Culture:
(81) Blood culture testing at Heidelberg University Hospital is routinely performed as described elsewhere (Gumbinger C et al. J Neurol Sci. 2013 325:46-50). Whole blood samples are obtained via direct venipuncture (e.g., antecubital vein) applying sterile techniques and 10 mL blood is inoculated to both an aerobic and an anaerobic liquid culture medium (BACTEC PLUS, BD Biosciences, Heidelberg, Germany). Cultures are incubated for 5 days (BACTEC, BD Biosciences, Heidelberg, Germany) and positive cultures are analyzed according to approved inhouse hospital standard techniques, including identification by VITEK2 (Biomerieux, Nuertingen, Germany) or MALDI TOF (Bruker, Madison, WI, USA) and automated antimicrobial susceptibility testing (VITEK 2).
(82) Culture-Based Diagnostic Procedures in Tracheal Secretion, Wound Swabs and Drainage Fluids:
(83) Briefly, tracheal aspirates and drainage fluids were streaked manually on Columbia (BD), chocolate (bM), MacConkey (bM), Schaedler and kanamycin-vancomycin (BD, Bi-plate) and chromogenic Candida agar (BD), while wound swabs were inoculated semi-automated by PREVI Isola instrument on the same agar types. All plates were incubated at 37 C. in 5% CO.sub.2 for 24 to 48h, except the Schaedler-KV bi-plates, which were incubated at 37 C. in an anaerobic chamber (GasPak; Becton, Dickinson, Franklin Lakes, NJ) for 48h as described (Mischnik A et al. J Clin Microbiol. 2012 50:2732-2736). Bacterial and fungal colonies were identified by MALDI-ToF mass spectrometry and automated AST was performed on VITEK II instruments (bM).
Group Definitions
(84) Candida spp. in the respiratory tract or in fluids from drainages were classified as colonization. Positive results in blood cultures, intraoperative swabs and Aspergillus spp. in deep respiratory tract specimens with accompanying pulmonary infiltrates were classified as infection.
(85) Anti-Candida-antibody titer: Candida albicans specific IgM, IgA and IgG antibodies in serum were detected and quantified using Serion ELISA Classic Candida albicans IgA/IgG/IgM (ESR 117A/G/M, Virion Serion, Wuerzburg, Germany) as described in the manufacturer's instructions using a Behring ELISA Processor (BEP III, Siemens Healthcare Diagnostics, Marburg, Germany), (Zou M et al. PLoS One. 2012 7:e43347).
Statistical Analyses
(86) The resulting data were entered into an electronic database (Excel 2010; Microsoft Corp, Redmond, USA) and evaluated using the SPSS software (Version 21.0; SPSS, Inc., Chicago, USA). Categorical data were summarized using absolute and relative frequencies. Quantitative data were summarized using median with quartiles. The Kolmogorov-Smirnov test was applied to check for normal distribution. Due to non-normally distributed data, non-parametric methods for evaluation were used (Chi-square test for categorical data, Mann-Whitney U test for continuous data). Appropriate cut-off values for the detection of a fungal infection were calculated using ROC analyses. A p-value <0.05 was considered statistically significant. Concerning symbolism and higher orders of significance: p<0.05: *, p<0.01: **, p<0.001: ***.
(87) Multiple comparison analysis has been performed by one-way analysis of variance (ANOVA) followed by a Dunnett's post hoc test.
Results
(88) Patient's characteristics. In total, 50 patients with septic shock were included in the presented investigation. Patients' characteristics are presented in Table 1. The underlying septic focus was the abdomen (n=43; 86%), followed by the lung (n=6; 12%), as well as the urogenital tract (n=1; 2%). The overall 28-day as well as 90-day mortality was 22% (n=11) and 34% (n=17), respectively. The median length of ICU as well as hospital stay was 20 days, and 44 days, respectively.
(89) TABLE-US-00001 TABLE 1 Patient's characteristics (n = 50) p for patients without with without fungal All fungal fungal isolates vs. patients isolates isolates patients with (n = 50) (n = 17) (n = 33) fungal isolates Gender male 38 (76) 11 (64.7) 27 (81.8) 0.160 Age (years) 66 (61-75) 71 (64-80) 66 (59-74) 0.117 BMI (kg/m.sup.2) 27.2 (24.4-30.9) 27.2 (25.7-34.9) 26.9 (23.1-30.9) 0.401 Postoperatively 31 9 (52.9) 22 (66.7) 0.206 peritonitis initial operation Kidney 2 (4) 0 (0) 2 (6.1) 0.431 Liver 11 (22) 1 (2.1) 10 (30.3) 0.047* Pancreas 2 (10) 1 (5.9) 1 (3.0) 0.569 GIT 38 ((76) 14 (82.4) 24 (72.7) 0.350 VAS 3 (6) 2 (11.8) 1 (3.0) 0.264 Others 12 (24) 3 (17.6) 9 (27.3) 0.350 48 h after hospital 25 (50) 7 (41.2) 18 (54.5) 0.276 admission NYHA 0-I 41 (82) 13 (76.4) 28 (84.8) 0.358 Diabetes mellitus 17 (34) 5 (29.4) 12 (36.3) 0.434 Arterial hypertension 34 (68) 12 (70.6) 22 (66.7) 0.520 Coronary heart 8 (16) 5 (29.4) 3 (9.1) 0.076 disease Chronic obstructive 10 (20) 5 (29.4) 5 (15.2) 0.204 lung disease Renal insufficiency 7 (14) 1 (5.9) 6 (18.2) 0.231 Renal replacement 15 (30) 2 (11.8) 13 (39.4) 0.041* therapy Liver cirrhosis 13 (26) 3 (17.6) 10 (30.3) 0.270 Oncological disease 33 (66) 11 (64.7) 22 (66.7) 0.566 APACHE II* 30 (28-35) 32 (28-36) 30 (28-34) 0.491 SOFA* 11 (10-14) 11 (10-14) 11 (10-14) 0.959 SAPS* 65 (49-75) 72 (48-75) 65 (51-72) 0.467 Candida colonization 22 (44) 0 (0) 22 (66.7) Candida infection 10 (20) 0 (0) 10 (30.3) Candidemia 3 (6) 0 (0) 3 (9.1) Aspergillus spp. 1 (3) 0 (0) 1 (3.0) Candida-Score 4 (4-4) 4 (4-4) 4 (4-4) 0.080 Duration of (hours) 145.5 (67.3-450) 89 (46-145) 181 (77-682) 0.015* mechanical ventilation ICU length of stay (days) 19.5 (12-44) 12 (3-17) 24 (15-46) 0.002** Hospital length of stay (days) 44 (23.3-68.5) 24 (12-40) 51 (39-78) 0.007** Tracheotomy 14 (28) 2 (11.8) 12 (36.3) 0.063 Anastomosis leakage 24 (48) 7 (41.2) 17 (51.5) 0.347 Fascia dehiscence 12 (24) 2 (11.8) 10 (30.3) 0.134 90 day mortality 17 (34) 8 (47.1) 9 (27.3) 0.175 28 day mortality 11 (22) 7 (41.2) 4 (12.1) 0.025*
(90) Data are presented as either number (with the corresponding percentage value) or median (with accompanying quartiles).
(91) The results of the cultured samples (standard diagnostics) has been used as criteria for the classification of patients in no fungal infection (n=17), (invasive) fungal infection (n=11) and fungal colonization (n=22) and were subdivided in different pathogens and locations of the infection, being presented in
(92) Fungal Pathogens and Infection Sites.
(93) Culture-based microbiological diagnostics: As assessed by culture-based microbiological diagnostics, fungal pathogens were present in 33 patients (66.0%), whereas 17 patients (34.0%) revealed negative fungal cultures. Fungal isolates were found in one or multiple locations in 25 (75.8%), or 8 (24.2%) patients respectively and were located at the following sites: respiratory tract (n=17; 51.5%), abdominal site (n=21; 63.6%) and blood culture (n=3; 9.1%). Characteristics of patients with or without fungal pathogens are presented in Table 1. Patients with fungal pathogens underwent more frequently liver surgery prior to study inclusion and the need for renal replacement therapy was shown to be significantly increased. Concerning further markers for morbidity, fungal-positive patients revealed a significant prolonged duration of mechanical ventilation and the need for tracheostomy tended to be increased. Moreover, length of ICU stay as well as hospital stay was significantly prolonged in patients with fungal pathogens. Surprisingly, 28-day mortality was significantly increased in patients without fungal pathogens, whereas 90-day mortality was shown to be comparable.
(94) Based on the group definitions as described in the methods section, colonization and infection was found in 22 (44.0%), and 11 (22.0%) patients, respectively. In colonized patients, 8 (16.0%) participants exclusively revealed Candida spp. in respiratory secretions (5 C. albicans, 1 C. albicans and glabrata, 2 C. albicans and C. spp), whereas in 6 (12.0%) patients Candida spp. could only be cultured from drainage fluids (3 C. albicans, 2 C. glabrata, 1 C. albicans and C. glabrata). Contrariwise, 8 (16.0%) patients were colonized at both sides (4 C. albicans, 1 C. albicans and C. spp., 3 C. albicans and C. glabrata). In infected patients, fungemia was found in 3 (6.0%) patients (2 C. albicans, 1 C. glabrata) and positive abdominal wound swabs were found in 7 (14.0%) patients (4 C. albicans, 1 C. glabrata, 1 C. krusei, 1 C. albicans and C. glabrata). Moreover, in one (2.0%) patient Aspergillus fumigatus was isolated in respiratory tract secretions. Concerning risk factors, liver surgery prior to study inclusion as well as liver cirrhosis could be observed more frequently in patients with a fungal infection. Moreover, the duration of ICU stay as well as mechanical ventilation was significantly prolonged and the need for tracheotomy was significantly increased in patients suffering from a fungal infection. Although morbidity was shown to be increased, mortality at 28 and 90 days did not differ significantly between infected and uninfected patients.
(95) Antifungal therapy. In total, 21 of 50 (42.0%) patients received an antifungal therapy during study participation. Of 17 patients without any fungal isolates, 2 (11.8%) patients received an empiric antifungal therapy. Of the remaining 33 patients with fungal isolates, 19 (57.6%) patients received an antifungal therapy, which was initiated in terms a specific therapy in 15 (78.9%) patients. Vice versa, treatment was initiated in terms of an empiric therapy in the remaining 4 (21.1%) cases, which was stopped later on in all of these patients. In 7 (33.3%) patients, the initial antifungal therapy was changed in the course of the disease.
(96) (1,3)--D-glucan (BG). Plasma concentrations of BG were comparable between the three subgroups throughout the entire study period and therefore failed to be of diagnostic value for the prediction of a fungal infection (data not shown). Even in patients suffering from candidemia, plasma concentrations of BG were not increased reliably.
(97) Galactomannan (GM). Plasma concentrations of GM remained below the cut-off value of <0.5 in 46 of 50 patients (92.0%). Contrariwise, 4 patients (8.0%) presented with sporadically increased plasma concentrations of GM above the cut-off value without any other (clinical, radiological, cultural) signs or risk factors for an IA (data not shown). In these cases, increased plasma concentrations of GM were most probably attributable to the underlying antibiotic therapy (e.g. piperacillin-tazobactam), which is well known to be associated with increased GM concentrations.
(98) One patient presented with the diagnosis of an IA as assessed by cultural detection of Aspergillus fumigatus in BALF, which was confirmed by high-resolution computed tomography. Moreover, GM concentrations in BALF were increased above the cut-off value, whereas plasma concentrations of GM remained below the cut-off value at all time points. Apart from septic shock as well as preexisting adipositas per magna and insulin-depending diabetes mellitus, the patient did not suffer from classical predisposing risk factors for IA (e.g. neutropenia, hemato-oncological diseases treated with cytotoxic agents, intake of corticosteroids, innate or acquired immunodeficiency). The patient was treated with liposomal amphotericin B for 6 weeks, which led to a decrease of GM in BALF below the cut-off value. Moreover, culture of BALF remained negative for Aspergillus fumigatus after the end the treatment period.
(99) Anti-Candida antibody titer. In the subgroup of patients without any fungal findings (n=17), 4 patients (23.5%) presented with a false positive anti-Candida antibody titer (>1:320), whereas colonized patients (n=22) were shown to have positive test results in 81.8% (n=18). Patients suffering from a fungal infection (n=11) also revealed positive test results in 81.8% (n=9), but unfortunately two patients presenting with candidemia (at sepsis onset) failed to show a positive anti-Candida antibody titer.
(100) TABLE-US-00002 TABLE 2 Biomarkers with significant values for the diagnosis and/or differentiation between invasive fungal infection and no invasive fungal infection. Fold change, 95% confidence interval (CI) and significant change, are indicated at different time points. Gene name T0 T1 T2 T3 T4 T5 T6 PIGR 1.9 2.8 2.1 (fold change/ 1.0-3.5 1.3-5.7 1.3-3.6 95% CI/ 0.046 0.004 0.003 p-value) ICAM1 1.6 1.6 1.6 1.9 1.7 (fold change/ 1.1-2.3 1.1-2.3 1.1-2.3 1.3-2.8 1.1-2.8 95% CI/ 0.013 0.023 0.009 <0.001 0.025 p-value) CPN1 0.6 0.7 0.4 (fold change/ 0.5-0.9 0.5-0.97 0.3-0.7 95% CI/ 0.005 0.03 0.001 p-value) HRG 0.4 0.2 (fold change/ 0.2-0.7 0.1-0.8 95% CI/ <0.001 0.016 p-value) THBS1 0.4 0.2 (fold change/ 0.2-0.98 0.04-0.8 95% CI/ 0.045 0.019 p-value) RAP1A 0.5 0.2 (fold change/ 0.2-0.95 0.04-0.7 95% CI/ 0.035 0.012 p-value) AHSG 0.4 0.3 (fold change/ 0.3-0.6 0.1-0.7 95% CI/ <0.001 0.004 p-value) FABP1 1.5 1.4 (fold change/ 1.03-2.1 1.007-1.96 95% CI/ 0.03 0.044 p-value) CT-proET-1 2.0 (fold change/ 1.1-3.5 95% CI/ 0.015 p-value) PCT 4.5 10.0 9.2 (fold change/ 1.7-11.7 2.5-40.9 1.2-69.0 95% CI/ 0.001 0.001 0.029 p-value)
(101) The Table 2 shows the results of the biomarkers PIGR, ICAM1, CPN1, HRG, THBS1, RAP1A AHSG, FABP1, ET-1 and PCT, tested at septic shock onset (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) with a significance (p value <0.05) and mean-fold change (presented below the p-value), whereby the significant biomarker values diagnose and/or differentiate invasive fungal infection compared to no invasive fungal infection. Biomarkers with mean-fold-changes below 1.0 indicate a downregulation of the biomarker (CPN1, THBS1, RAP1) and above 1.0 an upregulation of the biomarker (ICAM1, PIGR, FABP1, ET-1, PCT). Therefore the biomarkers show the same functionality in diagnosing and differentiation of invasive fungal infections vs. no invasive fungal infection and can be used alone or In combination (Table 3-6).
(102) TABLE-US-00003 TABLE 3 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with ICAM1 in all participating patients at sepsis onset (T0), and 1 day (T1), 2 days (T2) as well as 7 days (T3) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.739 0.727 0.156 0.01996 T1 0.790 0.727 0.156 0.02334 T2 0.818 0.727 0.063 0.0264 T3 0.841 0.727 0.094 0.0237
(103) Table 3: Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with ICAM1 in all participating patients at sepsis onset (T0), and 1 day (T1), 2 days (T2) as well as 7 days (T3) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis.
(104) TABLE-US-00004 TABLE 4 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with THBS1 in all participating patients at sepsis onset (T0), and 1 day (T1), 2 days (T2), 7 days (T3) as well as 14 days (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.279 0.30 0.7 0.1693 T1 0.350 0.3 0.708 0.1448 T2 0.287 0.30 0.7 0.1205 T3 0.387 0.1 0.333 0.3452 T4 0.129 0.2 0.917 0.2314
(105) Table 4 presents the diagnostic value of THBS for the diagnosis, differentiation, monitoring and prognosis/risk stratification of an (invasive) fungal infection in different time points, in patients with risk of having or getting a (invasive) fungal infection, in particular sepsis, especially septic shock. The results are transferrable to all kind of subjects, with and without special risk.
(106) TABLE-US-00005 TABLE 5 Area under the curve (AUC), sensitivity, 1-specificity and best cut- off from receiver operating characteristic (ROC) analysis with RAP1 (RAP1A/RAP1B/ RP1BL) in all participating patients at sepsis onset (T0), and 1 day (T1), 2 days (T2), days (T3) as well as 14 days (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.288 0.30 0.75 0.0479 T1 0.292 0.2 0.708 0.0462 T2 0.271 0.1 0.667 0.0476 T3 0.393 0.0 0.292 0.1022 T4 0.121 0.2 0.875 0.0618
(107) Table 5 presents the diagnostic value of RAP1 for the diagnosis, differentiation, monitoring and prognosis/risk stratification of an (invasive) fungal infection in different time points, in patients with risk of having or getting a (invasive) fungal infection, in particular sepsis, especially septic shock. The results are transferrable to all kind of subjects, with and without special risk.
(108) TABLE-US-00006 TABLE 6 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with VCL in all participating patients at sepsis onset (T0), and 1 day (T1), 2 days (T2), 7 days (T3) as well as 14 days (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.338 0.5 0.833 0.1533 T1 0.296 0.2 0.542 0.238 T2 0.258 0.1 0.625 0.213 T3 0.413 0.1 0.375 0.378 T4 0.150 0.3 0.917 0.2337
(109) Table 6 presents the diagnostic value of VCL for the diagnosis, differentiation, monitoring and prognosis/risk stratification of an (invasive) fungal infection in different time points, in patients with risk of having or getting a (invasive) fungal infection, in particular sepsis, especially septic shock. The results are transferrable to all kind of subjects, with and without special risk.
(110) TABLE-US-00007 TABLE 7 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with C-terminal Proendothelin-1 (CT-proET-1) in all participating patients at sepsis onset (T0), and 1 day (T1) afterwards with regard to the prediction of an invasive fungal infection (IFl) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Best Cut off Time point AUC Sensitivity 1-Specificity [pmol/l] T0 0.710 0.7 0.29 188.493 T1 0.716 0.8 0.323 155.492
(111) Table 7 presents the diagnostic value of CT-proET-1 for the diagnosis, differentiation, monitoring and prognosis/risk stratification of an (invasive) fungal infection in different time points, in patients with risk of having or getting a (invasive) fungal infection, in particular sepsis, especially septic shock. The results are transferrable to all kind of subjects, with and without special risk.
(112) TABLE-US-00008 TABLE 8 Biomarkers with significant values for the diagnosis and/or differentiation between (invasive) fungal infection and no invasive fungal infection or fungal colonization. Fold change, 95% confidence interval (CI) and significant change are indicated at different time points. Gene name T0 T1 T2 T3 T4 T5 T6 PIGR 1.9 2.0 2.5 2.1 (fold change/ 1.1-3.2 1.1-3.4 1.4-4.7 1.3-3.1 95% CI/ 0.023 0.012 0.002 0.001 p-value) ICAM1 1.5 1.5 1.7 1.8 1.7 (fold change/ 1.04-2.0 1.1-2.2 1.2-2.3 1.3-2.5 1.1-2.5 95% CI/ 0.027 0.009 0.001 <0.001 0.007 p-value) CPN1 0.7 0.7 0.6 (fold change/ 0.5-0.9 0.5-0.98 0.4-0.9 95% CI/ 0.01 0.036 0.013 p-value) HRG 0.5 0.3 (fold change/ 0.3-0.7 0.1-0.6 95% CI/ 0.001 0.001 p-value) THBS1 0.4 0.3 (fold change/ 0.2-0.8 0.1-0.7 95% CI/ 0.007 0.008 p-value) RAP1A 0.4 0.3 (fold change/ 0.2-0.7 0.1-0.7 95% CI/ 0.002 0.003 p-value) AHSG 0.5 0.4 (fold change/ 0.4-0.7 0.2-0.7 95% CI/ <0.001 0.002 p-value) VCL 0.6 0.4 0.4 (fold change/ 0.4-0.995 0.2-0.8 0.1-0.9 95% CI/ 0.047 0.003 0.019 p-value) PCT 4.6 8.9 7.1 7.2 (fold change/ 2.0-10.5 2.8-28.1 1.3-40.5 2.0-26.6 95% CI/ <0.001 <0.001 0.024 0.002 p-value)
(113) Table 8 shows the results of the biomarkers PIGR, ICAM1, CPN1, HRG, THBS1, RAP1A, AHSG, VCL and PCT, tested at septic shock onset (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) with a significance (p value <0.05) and mean-fold change (presented below the p-value), whereby the significant biomarker values diagnose and/or differentiate (invasive) fungal infection compared to no invasive fungal infection or fungal colonization. Biomarkers with Mean-fold-changes below 1.0 indicate a downregulation of the biomarker (PIGR, CPN1, HRG, THBS1, RAP1A (RAP1A/RAP1B/RAPBL), AHSG, VCL) and above 1.0 an upregulation of the biomarker (ICAM1, PCT). Therefore the biomarkers show the same functionality in diagnosing, and/or ruling out an invasive fungal infection and/or differentiation of an invasive fungal infection vs. no fungal infection or differentiation of an invasive fungal infection vs. no invasive fungal infection or fungal colonization and can be used alone or in combination.
(114) TABLE-US-00009 TABLE 9 Area under the curve (AUC) for prediction of an invasive fungal infection (IFI) compared to patients with fungal colonization at the time point of first fungal detection in microbiological samples. Marker AUC Sensitivity 1-Specificity Best Cut off ICAM1 0.707 0.727 0.273 0.02263 THBS1 0.302 0.273 0.7773 0.1713 RAP1 0.281 0.091 0.682 0.0544 VCL 0.264 0.091 0.773 0.2178
(115) TABLE-US-00010 TABLE 10 Area under the curve (AUC) for prediction of an invasive fungal infection (IFI) compared to patients with fungal colonization and patients without any fungal findings at the time point of first fungal detection in microbiological samples. In patients with no fungal findings, plasma concentrations of markers at sepsis onset were used. Marker AUC Sensitivity 1-Specificity Best Cut off ICAM1 0.767 0.727 0.154 0.02263 THBS1 0.336 0.091 0.513 0.23684 RAP1 0.322 0.091 0.615 0.05448 VCL 0.308 0.091 0.667 0.21783
(116) Table 9 and 10 present the predictive and/or diagnostic value of an (invasive) fungal infection and/or the ruling out of an (invasive) fungal infection (Tables 9 and 10) and/or the differentiation value of (invasive) fungal infection vs. fungal colonization (Table 9) of ICAM1, THBS1, RAP1 (RAP1A/RAP1B/RAPBL) and VCL and show the correlation with the first detection in microbiological samples. Therefore the biomarkers are usable for detecting the first onset of an (invasive) fungal infection and can differentiate between uncritical fungal colonization and a (invasive) fungal infection (Table 9).
(117) TABLE-US-00011 TABLE 11 Area under the curve (AUC), sensitivity, 1-specificity and best cut- off from receiver operating characteristic (ROC) analysis with ICAM1 in all participating patients change from 0 day (T0) to 1 day (T1), 0 day (T0) to 2 days (T2) or 0 day (T0) to 7 days (T3) after sepsis onset with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T1 vs T0 0.681 0.636 0.256 0.003245 T2 vs T0 0.741 0.727 0.205 0.00385 T3 vs T0 0.695 0.545 0.077 0.007489
(118) ROC analysis of ICAM1 change results in best diagnostic value of ICAM1 increase from T0 to T2 (target group: patients with an invasive fungal infection (IFI), controls: patients with a fungal colonization or without any fungal isolates).
(119) TABLE-US-00012 TABLE 12 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with PIGR in all participating patients 1 day (T1), 2 days (T2), 7 days (T3) as well as 14 days (T4) after sepsis onset with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T1 0.704 0.6 0.167 0.034 T2 0.729 0.5 0.042 0.0588 T3 0.846 0.9 0.292 0.0464 T4 0.833 0.8 0.208 0.0625
(120) TABLE-US-00013 TABLE 13 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with CPN1 in all participating patients at sepsis onset (T0), and 1 day (T1), 2 days (T2) as well as 7 days (T3) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.295 0.273 0.656 0.0428 T1 0.250 0.273 0.844 0.0379 T2 0.241 0.273 0.781 0.0483 T3 0.222 0.182 0.813 0.0358
(121) TABLE-US-00014 TABLE 14 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with HRG in all participating patients 14 days (T4) after sepsis onset with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T4 0.160 0.1 0.6 0.6408
(122) TABLE-US-00015 TABLE 15 Area under the curve (AUC), sensitivity, 1-specificity and best cut- off from receiver operating characteristic (ROC) analysis with AHSG in all participating patients 7 days (T3) and 14 days (T4) after sepsis onset with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T3 0.212 0.1 0.658 0.2664 T4 0.120 0.0 0.64 0.4125
(123) TABLE-US-00016 TABLE 16 Area under the curve (AUC), sensitivity, 1-specificity and best cut- off from receiver operating characteristic (ROC) analysis with FABP1 in all participating patients 1 day (T1) and 2 days (T2) after sepsis onset with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T1 0.716 0.818 0.333 0.0058 T2 0.837 1.0 0.303 0.0054
(124) A new and surprising finding was ICAM1 as biomarker for the diagnosis and/or risk prediction and/or risk stratification and/or monitoring and/or ruling in or ruling out of an invasive fungal infection, an fungal colonization and no fungal infection in a subject, in particular a risk group of getting or having an (invasive) fungal infection, especially sepsis e.g. septic shock.
(125)
(126) Plasma samples were collected at the onset of septic shock (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) afterwards and ICAM1 was measured.
(127)
(128)
(129)
(130)
(131) The
(132) ICAM1 increases on last time point compared to previous time point if patient died before 90 d, (
(133) A new and surprising finding was THBS1 as biomarker for the diagnosis and/or risk prediction and/or risk stratification and/or monitoring and/or ruling in or ruling out of an invasive fungal infection, an fungal colonization and no fungal infection in a subject, in particular a risk group of getting or having an (invasive) fungal infection, especially sepsis e.g. septic shock.
(134)
(135) Plasma samples were collected at the onset of septic shock (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) afterwards and THBS1 was measured.
(136)
(137) A new and surprising finding was RAP1 (RAP1A/RAP1B/RP1BL) as biomarker for the diagnosis and/or risk prediction and/or risk stratification and/or monitoring and/or ruling in or ruling out of an invasive fungal infection, an fungal colonization and no fungal infection in a subject, in particular a risk group of getting or having an (invasive) fungal infection, especially sepsis e.g. septic shock.
(138)
(139) Plasma samples were collected at the onset of septic shock (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) afterwards and RAP1 (RAP1A/RAP1B/RP1BL) was measured.
(140)
(141)
(142) TABLE-US-00017 TABLE 17 Biomarkers with significant values for the diagnosis and/or differentiation between (invasive) fungal infection and fungal colonization. Fold change, 95% confidence interval (CI) and significant change are indicated at different time points. Gene name T0 T1 T2 T3 T4 T5 T6 PIGR 2.1 2.1 (fold change/ 1.1-4.0 1.4-3.4 95% CI/ 0.021 0.001 p-value) ICAM1 1.5 1.5 1.6 1.6 1.6 (fold change/ 1.02-2.1 1.03-2.1 1.1-2.3 1.1-2.2 1.1-2.5 95% CI/ 0.037 0.029 0.006 0.005 0.015 p-value) HRG 0.5 0.4 0.3 (fold change/ 0.3-0.8 0.2-0.9 0.1-0.6 95% CI/ 0.004 0.03 0.001 p-value) THBS1 0.4 0.4 (fold change/ 0.2-0.9 0.1-0.95 95% CI/ 0.022 0.038 p-value) RAP1A 0.4 0.3 (fold change/ 0.2-0.8 0.1-0.8 95% CI/ 0.004 0.015 p-value) AHSG 0.6 0.5 0.4 (fold change/ 0.4-0.8 0.3-0.9 0.2-0.8 95% CI/ 0.002 0.019 0.003 p-value) VCL 0.6 0.6 0.4 (fold change/ 0.3-0.98 0.3-0.97 0.2-0.8 95% CI/ 0.041 0.035 0.007 p-value) FABP1 1.4 1.4 (fold change/ 1.003-1.9 1.1-1.9 95% CI/ 0.047 0.009 p-value) PCT 3.6 7.1 8.7 8.0 (fold change/ 1.5-8.7 2.1-24.0 1.5-50.7 2.2-29.4 95% CI/ 0.002 0.001 0.014 0.002 p-value)
(143) Table 17 shows the results of the biomarkers PIGR, ICAM1, HRG, THBS1, RAP1A AHSG, VCL FABP1 and PCT, tested at septic shock onset (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) with a significance (p value <0.05) and mean-fold change (presented below the p-value), whereby the significant biomarker values diagnose and/or differentiate (invasive) fungal infection compared to fungal colonization. Biomarkers with fold-changes below 1.0 indicate a downregulation of the biomarker (HRG, THBS1, RAP1A (RAP1A/RAP1B/RAPBL), AHSG, VCL) and above 1.0 an upregulation of the biomarker (ICAM1, FABP1, PCT). Therefore the biomarkers show the same functionality in diagnosing, and/or ruling out an invasive fungal infection and/or differentiation of an invasive fungal infection vs. no invasive fungal infection and/or differentiation of an invasive fungal infection vs. no invasive fungal infection and/or fungal colonization and differentiation of an invasive fungal infection vs. fungal colonization and can be used alone or in combination.
(144)
(145)
(146) A new and surprising finding was VCL as biomarker for the diagnosis and/or risk prediction and/or risk stratification and/or monitoring and/or ruling in or ruling out of an invasive fungal infection, an fungal colonization and no fungal infection in a subject, in particular a risk group of getting or having an (invasive) fungal infection, especially sepsis e.g. septic shock.
(147)
(148) Plasma samples were collected at the onset of septic shock (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) afterwards and VCL was measured.
(149)
(150)
(151)
(152)
(153) A new and surprising finding was CT-proET-1 as biomarker for the diagnosis and/or risk prediction and/or risk stratification and/or monitoring and/or ruling in or ruling out of an invasive fungal infection, an fungal colonization and no fungal infection in a subject, in particular a risk group of getting or having an (invasive) fungal infection, especially sepsis e.g. septic shock.
(154)
(155) Plasma samples were collected at the onset of septic shock (T0), and 1 day (T1) afterwards and CT-proET-1 was measured.
(156)
(157) At the time point of a first fungal detection, ICAM1, THBS1, RAP1A; VCL and CT-proET-1 are able to differentiate invasive fungal infection from fungal colonization and patients without fungal findings (
(158)
(159) In addition, plasma levels of sICAM-1 were assessed by Human sICAM-1 Platinum ELISA (eBioscience, Thermo Fisher Scientific), an immunoassay-based procedure, for the time points T0 as well as T1.
(160) TABLE-US-00018 TABLE 18 Area under the curve (AUC), sensitivity with 95%-confidence intervals (CI), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with sICAM-1 in all participating patients at sepsis onset (T0) and 1 day (T1) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis. AUC (with Best Cut off Time point 95%-CI) Sensitivity 1-Specificity (ng/L) T0 0.656 0.545 0.121 1705 (438-0.874) T1 0.716 0.727 0.182 1591 (0.502-0.931)
(161)
(162) (A) Plasma concentrations of sICAM-1 were measured in patients suffering from septic shock with an invasive fungal infection (IFI, dark grey box), a fungal colonization (light grey box) or without any fungal findings (white box). Plasma samples were collected at the onset of septic shock (T0) and 1 day (T1) afterwards. Data in box plots are given as median, 25th percentile, 75th percentile with the 10th as well as 90th percentile at the end of the whiskers. Concerning symbolism and higher orders of significance: p<0.05: *.
(163) (B) Receiver operating characteristic (ROC) analysis with sICAM-1 in all participating patients at sepsis onset (T0) and 1 day (T1) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(164) TABLE-US-00019 TABLE 19 Receiver Operator Curve (ROC)-analyses for the measurement of PCT. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound PCT T0 .525 .105 .821 .319 .731 PCT T1 .533 .112 .762 .314 .753 PCT T2 .583 .109 .450 .370 .796 PCT T3 .879 .067 .001 .759 .999 PCT T4 .896 .055 .000 .789 1.000 .sup.aUnder the nonparametric assumption .sup.bNull hypothesis: true area = .05
(165)
(166) Receiver operating characteristic (ROC) analysis with PCT in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(167) TABLE-US-00020 TABLE 20 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT and ICAM-1. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .729 .109 .038 .516 .942 probability Predicted .779 .110 .011 .564 .995 probability Predicted .813 .107 .005 .602 1.000 probability Predicted .900 .080 .000 .743 1.000 probability Predicted .837 .070 .002 .700 .975 probability
(168)
(169) Receiver operating characteristic (ROC) analysis with PCT and ICAM in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(170) TABLE-US-00021 TABLE 21 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT, ICAM-1 and ADM. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .796 .096 .008 .607 .984 probability Predicted .813 .096 .005 .625 1.000 probability Predicted .830 .103 .003 .628 1.000 probability Predicted .909 .072 .000 .768 1.000 probability Predicted .896 .056 .000 .785 1.000 probability
(171)
(172) Receiver operating characteristic (ROC) analysis with PCT, ICAM and ADM in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(173) TABLE-US-00022 TABLE 22 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT, ICAM-1, ADM and IL17. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .803 .104 .009 .600 1.000 probability Predicted .838 .098 .004 .645 1.000 probability Predicted .803 .118 .009 .573 1.000 probability Predicted .909 .074 .000 .764 1.000 probability Predicted .919 .053 .000 .813 1.000 probability
(174)
(175) Receiver operating characteristic (ROC) analysis with PCT, ICAM-1, ADM and IL17 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(176) TABLE-US-00023 TABLE 23 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT and ADM. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .674 .107 .117 .464 .884 probability Predicted .661 .101 .147 .463 .858 probability Predicted .683 .106 .100 .475 .890 probability Predicted .883 .064 .001 .756 1.000 probability Predicted .887 .059 .000 .772 1.000 probability
(177)
(178) Receiver operating characteristic (ROC) analysis with PCT and ADM in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(179) TABLE-US-00024 TABLE 24 Receiver Operator Curve (ROC)-analyses for the combined measurement of ADM and ICAM-1. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .787 .087 .010 .617 .957 probability Predicted .835 .080 .003 .677 .992 probability Predicted .852 .091 .002 .673 1.000 probability Predicted .887 .071 .000 .748 1.000 probability Predicted .909 .052 .000 .808 1.000 probability
(180)
(181) Receiver operating characteristic (ROC) analysis with ADM and ICAM-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(182) TABLE-US-00025 TABLE 25 Receiver Operator Curve (ROC)-analyses for the combined measurement of ADM, ICAM-1 and IL17. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .788 .100 .013 .593 .983 probability Predicted .848 .086 .003 .679 1.000 probability Predicted .828 .102 .005 .628 1.000 probability Predicted .879 .077 .001 .727 1.000 probability Predicted .899 .067 .001 .767 1.000 probability .sup.bNull hypothese: true area = 0.5
(183)
(184) Receiver operating characteristic (ROC) analysis with ADM, ICAM and IL17 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(185) TABLE-US-00026 TABLE 26 Receiver Operator Curve (ROC)-analyses for the measurement of ADM. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound T0 .700 .097 .070 .509 .891 T1 .663 .101 .140 .464 .861 T2 .725 .100 .041 .529 .921 T3 .783 .093 .010 .600 .966 T4 .908 .051 .000 .808 1.000 .sup.aUnder the nonparametric assumption .sup.bNull hypothesis: true area = .05
(186)
(187) Receiver operating characteristic (ROC) analysis with ADM in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(188) TABLE-US-00027 TABLE 27 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT and THBS-1. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .687 .099 .089 .493 .882 probability Predicted .629 .116 .241 .402 .857 probability Predicted .708 .102 .059 .509 .908 probability Predicted .879 .061 .001 .759 .999 probability Predicted .879 .067 .001 .747 1.000 probability
(189)
(190) Receiver operating characteristic (ROC) analysis with PCT and THBS-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(191) TABLE-US-00028 TABLE 28 Receiver Operator Curve (ROC)-analyses for the combined measurement of ADM and THBS-1. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .730 .102 .038 .531 .930 probability Predicted .704 .112 .066 .485 .923 probability Predicted .735 .102 .034 .534 .965 probability Predicted .804 .079 .006 .649 .960 probability Predicted .909 .052 .000 .807 1.000 probability
(192)
(193) Receiver operating characteristic (ROC) analysis with ADM and THBS-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(194) TABLE-US-00029 TABLE 29 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT, ADM and THBS-1. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .709 .106 .060 .501 .917 probability Predicted .657 .111 .158 .440 .874 probability Predicted .726 .104 .042 .522 .930 probability Predicted .870 .067 .001 .739 1.000 probability Predicted .922 .047 .000 .830 1.000 probability
(195)
(196) Receiver operating characteristic (ROC) analysis with PCT, ADM and THBS-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(197) TABLE-US-00030 TABLE 30 Receiver Operator Curve (ROC)-analyses for the combined measurement of PCT and VCL. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .638 .103 .212 .436 .839 probability Predicted .700 .104 .070 .496 .904 probability Predicted .742 .094 .028 .557 .927 probability Predicted .875 .063 .001 .752 .998 probability Predicted .862 .064 .001 .736 .989 probability
(198)
(199) Receiver operating characteristic (ROC) analysis with PCT and VCL in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(200) TABLE-US-00031 TABLE 31 Receiver Operator Curve (ROC)-analyses for the combined measurement of ADM and VCL. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .709 .106 .060 .500 .917 probability Predicted .757 .103 .021 .554 .959 probability Predicted .730 .098 .038 .539 .922 probability Predicted .765 .091 .017 .587 .943 probability Predicted .917 .047 .000 .826 1.000 probability
(201)
(202) Receiver operating characteristic (ROC) analysis with ADM and VCL in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(203) TABLE-US-00032 TABLE 32 Receiver Operator Curve (ROC)-analyses for the combined measurement of ADM, VCL and PCT. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .696 .106 .078 .488 .903 probability Predicted .739 .100 .031 .543 .935 probability Predicted .748 .097 .026 .558 .937 probability Predicted .878 .066 .001 .748 1.000 probability Predicted .922 .046 .000 .831 1.000 probability
(204)
(205) Receiver operating characteristic (ROC) analysis with ADM, VCL and PCT in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(206) TABLE-US-00033 TABLE 33 Receiver Operator Curve (ROC)-analyses for the combined measurement of ICAM1 and THBS-1. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .787 .091 .009 .609 .966 probability Predicted .808 .094 .005 .625 .992 probability Predicted .833 .090 .002 .656 1.000 probability Predicted .887 .081 .000 .730 1.000 probability Predicted .887 .058 .000 .775 1.000 probability
(207)
(208) Receiver operating characteristic (ROC) analysis with ICAM1 and THBS-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(209) TABLE-US-00034 TABLE 34 Receiver Operator Curve (ROC)-analyses for the combined measurement of ICAM1 and VCL. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .771 .098 .014 .579 .962 probability Predicted .837 .078 .002 .684 .991 probability Predicted .883 .075 .001 .736 1.000 probability Predicted .854 .086 .001 .686 1.000 probability Predicted .883 .061 .001 .763 1.000 probability
(210)
(211) Receiver operating characteristic (ROC) analysis with ICAM1 and VCL in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(212) TABLE-US-00035 TABLE 35 Receiver Operator Curve (ROC)-analyses for the combined measurement of ICAM1, THBS-1 and VCL. ROC-analyses for fungally infected vs. fungally colonized or patients without any fungal findings. Data are given as AUCs with 95%-confidence intervals (CI) or absolute values for sensitivity and specificity. Abbreviations: AUC, area under the curve; CI, confidence interval. Area Under the Curve Asymptotic 95% Confidence Interval Test Result Std. Asymptotic Lower Upper Variable(s) Area Error.sup.a Sig..sup.b Bound Bound Predicted .800 .087 .007 .629 .971 probability Predicted .846 .076 .002 .696 .996 probability Predicted .896 .068 .000 .763 1.000 probability Predicted .921 .046 .000 .831 1.000 probability Predicted .896 .057 .000 .785 1.000 probability
(213)
(214) Receiver operating characteristic (ROC) analysis with ICAM1, THBS-1 and VCL in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(215)
(216)
(217) TABLE-US-00036 TABLE 37 Area under the curve (AUC), sensitivity with 95%-confidence intervals (CI), sensitivity and 1-specificity from receiver operating characteristic (ROC) analysis with sICAM-1, thrombospondin-1 and vinculin in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis. AUC (with Time point 95%-CI) Sensitivity 1-Specificity T0 0.800 0.700 0.174 (0.629-0.917) T1 0.846 0.800 0.174 (696-0.996) T2 0.896 0.700 0.00 (0.763-1.00) T3 0.921 0.900 0.174 (0.831-1.0) T4 0.896 0.800 0.130 (0.785-1.00)
(218)
(219) Receiver operating characteristic (ROC) analysis with sICAM-1, thrombospondin-1 and vinculin in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
(220) TABLE-US-00037 TABLE 38 Area under the curve (AUC), sensitivity with 95%-confidence intervals (CI), sensitivity and 1-specificity from receiver operating characteristic (ROC) analysis with MR-proADM and sICAM-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis. AUC (with Time point 95%-CI) Sensitivity 1-Specificity T0 0.787 0.700 0.083 (0.617-0.957) T1 0.835 0.800 0.250 (0.677-0.992) T2 0.852 0.900 0.208 (0.673-1.000) T3 0.887 1.000 0.292 (0.748-1.000) T4 0.909 0.900 0.208 (0.808-1.000)
(221)
(222) Receiver operating characteristic (ROC) analysis with MR-proADM and sICAM-1 in all participating patients at sepsis onset (T0), day 1 (T1), day 2 (T2), day 7 (T3) and 14 day (T4) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC-analysis.
Example 2: IFI in the Context of Liver Transplantation, Especially Patients Following Liver Transplantation
(223) The following experiments were performed as described in Example 1. In brief, plasma concentrations of ICAM1, MR-proADM or ICAM1 and MR-proADM were measured in patients following liver transplantation (LTX) with an invasive fungal infection, a fungal colonization or without any fungal findings. In total, 93 patients following LTX were screened for the emergence of IFIs by the use of culture-based as well as image-producing procedures. In parallel, plasma samples were collected on day of liver transplantation (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4), 21 days (T5) and 28 days (T6) afterwards.
(224) 2.1 ICAM1 for the Detection of an Invasive Fungal Infection Following Liver Transplantation
(225) TABLE-US-00038 TABLE 39 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with ICAM1 in all participating patients on day of liver transplantation (T0), and 1 day (T1), 14 days (T4) as well as 21 days (T5) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. AUC (with Time point 95%-CI) Sensitivity 1-Specificity Best Cut off T0 0.574 0.5 0.238 0.022375364 T1 0.533 0.625 0.476 0.023814595 T4 0.714 0.875 0.429 0.025729737 (0.546-0.882) T5 0.783 0.875 0.357 0.024646877 (0.645-0.920)
2.2 ICAM1 for the Detection of Fungal Pathogens in IFI Vs. Fungal Colonization Following Liver Transplantation
(226) TABLE-US-00039 TABLE 40 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with ICAM1 in patients with an invasive fungal infection (IFI) or fungal colonization on day of liver transplantation (T0), 1 day (T1) and 2 days (T2) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an IFI represented the target group, whereas patients with a fungal colonization served as controls for this ROC analysis. Data of patients without any fungal findings were not included in this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.708 0.5 0.0 0.025947727 T1 0.833 0.75 0.0 0.022376599 T2 0.917 0.75 0.0 0.019209418
2.2 MR-proADM for the Detection of Fungal Pathogens in IFI Vs. Fungal Colonization Following Liver Transplantation
(227) TABLE-US-00040 TABLE 41 Area under the curve (AUC), sensitivity, 1-specificity and best cut-off from receiver operating characteristic (ROC) analysis with MR-proADM in patients with an invasive fungal infection (IFI) or fungal colonization on day of liver transplantation (T0), 1 day (T1) and 2 days (T2) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an IFI represented the target group, whereas patients with a fungal colonization served as controls for this ROC analysis. Data of patients without any fungal findings were not included in this ROC analysis. Time point AUC Sensitivity 1-Specificity Best Cut off T0 0.679 1.0 0.5 3.06 T1 0.750 1.0 0.5 4.69 T2 0.857 0.857 0.25 5.80
(228)
(229) 2.3 ICAM-1 and MR-proADM for the Detection of an Invasive Fungal Infection Following Liver Transplantation
(230) TABLE-US-00041 TABLE 42 Area under the curve (AUC), sensitivity and 1-specificity from receiver operating characteristic (ROC) analysis with ICAM1 and MR-proADM in all participating patients on day of liver transplantation (T0), and 1 day (T1), 2 days (T2), 7 days (T3), 14 days (T4) as well as 21 days (T5) afterwards with regard to the prediction of an invasive fungal infection (IFI) up to day 28. Patients suffering from an invasive fungal infection (IFI) represented the target group, whereas both, patients with a fungal colonization as well as patients without any fungal isolates served as controls for this ROC analysis. AUC (with Time point 95%-CI) Sensitivity 1-Specificity T0 0.741 0.571 0.0 (0.465-1.00) T1 0.511 0.286 0.0 T2 0.575 0.429 0.105 T3 0.865 0.857 0.105 (0.706-1.00) T4 0.898 0.857 0.053 (0.758-1.00) T5 0.902 0.857 0.079
(231) TABLE-US-00042 AMINOACIDSEQUENCES Marker Aminoacidsequence(SEQIDNO) Intercellular MAPSSPRPALPALLVLLGALFPGPGNAQTS adhesion VSPSKVILPRGGSVLVTCSTSCDQPKLLGI molecule1 ETPLPKKELLLPGNNRKVYELSNVQEDSQP (ICAM1) MCYSNCPDGQSTAKTFLTVYWTPERVELAP UniprotNo.: LPSWQPVGKNLTLRCQVEGGAPRANLTVVL P05362 LRGEKELKREPAVGEPAEVTTTVLVRRDHH Length: GANFSCRTELDLRPQGLELFENTSAPYQLQ 532aa TFVLPATPPQLVSPRVLEVDTQGTVVCSLD GLFPVSEAQVHLALGDQRLNPTVTYGNDSF SAKASVSVTAEDEGTQRLTCAVILGNQSQE TLQTVTIYSFPAPNVILTKPEVSEGTEVTV KCEAHPRAKVTLNGVPAQPLGPRAQLLLKA TPEDNGRSFSCSATLEVAGQLIHKNQTREL RVLYGPRLDERDCPGNWTWPENSQQTPMCQ AWGNPLPELKCLKDGTFPLPIGESVTVTRD LEGTYLCRARSTQGEVTRKVTVNVLSPRYE IVIITVVAAAVIMGTAGLSTYLYNRQRKIK KYRLQQAQKGTPMKPNTQATPP (SEQIDNO1) Alpha-2-HS- MKSLVLLLCLAQLWGCHSAPHGPGLIYRQP glycoprotein NCDDPETEEAALVAIDYINQNLPWGYKHTL (AHSG) NQIDEVKVWPQQPSGELFEIEIDTLETTCH UniprotNo.: VLDPTPVARCSVRQLKEHAVEGDCDFQLLK P02765 LDGKFSVVYAKCDSSPDSAEDVRKVCQDCP Length: LLAPLNDTRVVHAAKAALAAFNAQNNGSNF 367aa QLEEISRAQLVPLPPSTYVEFTVSGTDCVA KEATEAAKCNLLAEKQYGFCKATLSEKLGG AEVAVTCTVFQTQPVTSQPQPEGANEAVPT PVVDPDAPPSPPLGAPGLPPAGSPPDSHVL LAAPPGHQLHRAHYDLRHTFMGVVSLGSPS GEVSHPRKTRTVVQPSVGAAAGPVVPPCPG RIRHFKV (SEQIDNO2) Carboxypeptidase MSDLLSVFLHLLLLFKLVAPVTFRHHRYDD Ncatalytic LVRTLYKVQNECPGITRVYSIGRSVEGRHL chain YVLEFSDHPGIHEPLEPEVKYVGNMHGNEA (CPN1) LGRELMLQLSEFLCEEFRNRNQRIVQLIQD UniprotNo.: TRIHILPSMNPDGYEVAAAQGPNKPGYLVG P15169 RNNANGVDLNRNFPDLNTYIYYNEKYGGPN Length: HHLPLPDNWKSQVEPETRAVIRWMHSFNFV 458aa LSANLHGGAVVANYPYDKSFEHRVRGVRRT ASTPTPDDKLFQKLAKVYSYAHGWMFQGWN CGDYFPDGITNGASWYSLSKGMQDFNYLHT NCFEITLELSCDKFPPEEELQREWLGNREA LIQFLEQVHQGIKGMVLDENYNNLANAVIS VSGINHDVTSGDHGDYFRLLLPGIYTVSAT APGYDPETVTVTVGPAEPTLVNFHLKRSIP QVSPVRRAPSRRHGVRAKVQPQARKKEMEM RQLQRGPA (SEQIDNO3) Fattyacid- MSFSGKYQLQSQENFEAFMKAIGLPEELIQ binding KCKDIKGVSEIVQNGKHFKFTITAGSKVIQ protein NEFTVGEECELETMTGEKVKTVVQLEGDNK (FABP1) LVTTFKNIKSVTELNGDIITNTMTLGDIVF UniprotNo.: KRISKRI P07148 (SEQIDNO4) Length: 127aa Histidine-rich MKALIAALLLITLQYSCAVSPTDCSAVEPE glycoprotein AEKALDLINKRRRDGYLFQLLRIADAHLDR (HRG) VENTTVYYLVLDVQESDCSVLSRKYWNDCE UniprotNo.: PPDSRRPSEIVIGQCKVIATRHSHESQDLR P04196 VIDFNCTTSSVSSALANTKDSPVLIDFFED Length: TERYRKQANKALEKYKEENDDFASFRVDRI 525aa ERVARVRGGEGTGYFVDFSVRNCPRHHFPR HPNVFGFCRADLFYDVEALDLESPKNLVIN CEVFDPQEHENINGVPPHLGHPFHWGGHER SSTTKPPFKPHGSRDHHHPHKPHEHGPPPP PDERDHSHGPPLPQGPPPLLPMSCSSCQHA TFGTNGAQRHSHNNNSSDLHPHKHHSHEQH PHGHHPHAHHPHEHDTHRQHPHGHHPHGHH PHGHHPHGHHPHGHHPHCHDFQDYGPCDPP PHNQGHCCHGHGPPPGHLRRRGPGKGPRPF HCRQIGSVYRLPPLRKGEVLPLPEANFPSF PLPHHKHPLKPDNQPFPQSVSESCPGKFKS GFPQVSMFFTHTFPK (SEQIDNO5) Polymeric MLLFVLTCLLAVFPAISTKSPIFGPEEVNS immunoglobulin VEGNSVSITCYYPPTSVNRHTRKYWCRQGA receptor RGGCITLISSEGYVSSKYAGRANLTNFPEN (PIGR) GTFVVNIAQLSQDDSGRYKCGLGINSRGLS UniprotNo.: FDVSLEVSQGPGLLNDTKVYTVDLGRTVTI P01833 NCPFKTENAQKRKSLYKQIGLYPVLVIDSS Length: GYVNPNYTGRIRLDIQGTGQLLFSVVINQL 764aa RLSDAGQYLCQAGDDSNSNKKNADLQVLKP EPELVYEDLRGSVTFHCALGPEVANVAKFL CRQSSGENCDVVVNTLGKRAPAFEGRILLN PQDKDGSFSVVITGLRKEDAGRYLCGAHSD GQLQEGSPIQAWQLFVNEESTIPRSPTVVK GVAGGSVAVLCPYNRKESKSIKYWCLWEGA QNGRCPLLVDSEGWVKAQYEGRLSLLEEPG NGTFTVILNQLTSRDAGFYWCLTNGDTLWR TTVEIKIIEGEPNLKVPGNVTAVLGETLKV PCHFPCKFSSYEKYWCKWNNTGCQALPSQD EGPSKAFVNCDENSRLVSLTLNLVTRADEG WYWCGVKQGHFYGETAAVYVAVEERKAAGS RDVSLAKADAAPDEKVLDSGFREIENKAIQ DPRLFAEEKAVADTRDQADGSRASVDSGSS EEQGGSSRALVSTLVPLGLVLAVGAVAVGV ARARHRKNVDRVSIRSYRTDISMSDFENSR EFGANDNMGASSITQETSLGGKEEFVATTE STTETKEPKKAKRSSKEEAEMAYKDFLLQS STVAAEAQDGPQEA (SEQIDNO6) Ras-related MREYKLVVLGSGGVGKSALTVQFVQGIFVE protein KYDPTIEDSYRKQVEVDCQQCMLEILDTAG Rap-1A TEQFTAMRDLYMKNGQGFALVYSITAQSTF (RAP1A) NDLQDLREQILRVKDTEDVPMILVGNKCDL UniprotNo.: EDERVVGKEQGQNLARQWCNCAFLESSAKS P62834 KINVNEIFYDLVRQINRKTPVEKKKPKKKS Length: CLLL 184aa (SEQIDNO7) Ras-related MREYKLVVLGSGGVGKSALTVQFVQGIFVE protein KYDPTIEDSYRKQVEVDAQQCMLEILDTAG Rap-1b TEQFTAMRDLYMKNGQGFALVYSITAQSTF (RAP1B) NDLQDLREQILRVKDTDDVPMILVGNKCDL UniprotNo.: EDERVVGKEQGQNLARQWNNCAFLESSAKS P61224 KINVNEIFYDLVRQINRKTPVPGKARKKSS isoform1 CQLL(SEQIDNO8) Length: 184aa Ras-related MREYKLVVLGSRGVGKSALTVQFVQGIFVE protein KYDPTIEDSYREQVEVDAQQCMLEILDTAG Rap-1b- TEQFTAMRDLYMKNGQGFALVYSITAQSTF likeprotein NDLQDLREQILRVKDTDDVPMILVGNKCDL (RP1BL) EDERVVGKEQGQNLARQWNNCAFLESSAKS UniprotNo.: KINVNEIFYDLVRQINRKTPVPGKARKKSS A6NIZ1 CQLL Length: (SEQIDNO9) 184aa Thrombospondin-1 a)Isoform1: (THBS1) MGLAWGLGVLFLMHVCGTNRIPESGGDNSV UniprotNo.: FDIFELTGAARKGSGRRLVKGPDPSSPAFR P07996 IEDANLIPPVPDDKFQDLVDAVRAEKGFLL Length: LASLRQMKKTRGTLLALERKDHSGQVFSVV a)isoform1: SNGKAGTLDLSLTVQGKQHVVSVEEALLAT 1,170aa GQWKSITLFVQEDRAQLYIDCEKMENAELD b)isoform2: VPIQSVFTRDLASIARLRIAKGGVNDNFQG 1,085aa VLQNVRFVFGTTPEDILRNKGCSSSTSVLL TLDNNVVNGSSPAIRTNYIGHKTKDLQAIC GISCDELSSMVLELRGLRTIVTTLQDSIRK VTEENKELANELRRPPLCYHNGVQYRNNEE WTVDSCTECHCQNSVTICKKVSCPIMPCSN ATVPDGECCPRCWPSDSADDGWSPWSEWTS CSTSCGNGIQQRGRSCDSLNNRCEGSSVQT RTCHIQECDKRFKQDGGWSHWSPWSSCSVT CGDGVITRIRLCNSPSPQMNGKPCEGEARE TKACKKDACPINGGWGPWSPWDICSVTCGG GVQKRSRLCNNPTPQFGGKDCVGDVTENQI CNKQDCPIDGCLSNPCFAGVKCTSYPDGSW KCGACPPGYSGNGIQCTDVDECKEVPDACF NHNGEHRCENTDPGYNCLPCPPRFTGSQPF GQGVEHATANKQVCKPRNPCTDGTHDCNKN AKCNYLGHYSDPMYRCECKPGYAGNGIICG EDTDLDGWPNENLVCVANATYHCKKDNCPN LPNSGQEDYDKDGIGDACDDDDDNDKIPDD RDNCPFHYNPAQYDYDRDDVGDRCDNCPYN HNPDQADTDNNGEGDACAADIDGDGILNER DNCQYVYNVDQRDTDMDGVGDQCDNCPLEH NPDQLDSDSDRIGDTCDNNQDIDEDGHQNN LDNCPYVPNANQADHDKDGKGDACDHDDDN DGIPDDKDNCRLVPNPDQKDSDGDGRGDAC KDDFDHDSVPDIDDICPENVDISETDFRRF QMIPLDPKGTSQNDPNWVVRHQGKELVQTV NCDPGLAVGYDEFNAVDFSGTFFINTERDD DYAGFVFGYQSSSRFYVVMWKQVTQSYWDT NPTRAQGYSGLSVKVVNSTTGPGEHLRNAL WHTGNTPGQVRTLWHDPRHIGWKDFTAYRW RLSHRPKTGFIRVVMYEGKKIMADSGPIYD KTYAGGRLGLFVFSQEMVFFSDLKYECRDP (SEQIDNO10) b)Isoform2: MGLAWGLGVLFLMHVCGTLLALERKDHSGQ VFSVVSNGKAGTLDLSLTVQGKQHVVSVEE ALLATGQWKSITLFVQEDRAQLYIDCEKME NAELDVPIQSVFTRDLASIARLRIAKGGVN DNFQGVLQNVRFVFGTTPEDILRNKGCSSS TSVLLTLDNNVVNGSSPAIRTNYIGHKTKD LQAICGISCDELSSMVLELRGLRTIVTTLQ DSIRKVTEENKELANELRRPPLCYHNGVQY RNNEEWTVDSCTECHCQNSVTICKKVSCPI MPCSNATVPDGECCPRCWPSDSADDGWSPW SEWTSCSTSCGNGIQQRGRSCDSLNNRCEG SSVQTRTCHIQECDKRFKQDGGWSHWSPWS SCSVTCGDGVITRIRLCNSPSPQMNGKPCE GEARETKACKKDACPINGGWGPWSPWDICS VTCGGGVQKRSRLCNNPTPQFGGKDCVGDV TENQICNKQDCPIDGCLSNPCFAGVKCTSY PDGSWKCGACPPGYSGNGIQCTDVDECKEV PDACFNHNGEHRCENTDPGYNCLPCPPRFT GSQPFGQGVEHATANKQVCKPRNPCTDGTH DCNKNAKCNYLGHYSDPMYRCECKPGYAGN GIICGEDTDLDGWPNENLVCVANATYHCKK DNCPNLPNSGQEDYDKDGIGDACDDDDDND KIPDDRDNCPFHYNPAQYDYDRDDVGDRCD NCPYNHNPDQADTDNNGEGDACAADIDGDG ILNERDNCQYVYNVDQRDTDMDGVGDQCDN CPLEHNPDQLDSDSDRIGDTCDNNQDIDED GHQNNLDNCPYVPNANQADHDKDGKGDACD HDDDNDGIPDDKDNCRLVPNPDQKDSDGDG RGDACKDDFDHDSVPDIDDICPENVDISET DFRRFQMIPLDPKGTSQNDPNWVVRHQGKE LVQTVNCDPGLAVGYDEFNAVDFSGTFFIN TERDDDYAGFVFGYQSSSRFYVVMWKQVTQ SYWDTNPTRAQGYSGLSVKVVNSTTGPGEH LRNALWHTGNTPGQVRTLWHDPRHIGWKDF TAYRWRLSHRPKTGFIRVVMYEGKKIMADS GPIYDKTYAGGRLGLFVFSQEMVFFSDLKY ECRDP (SEQIDNO11) Vinculin a)Isoform1: (VCL) MPVFHTRTIESILEPVAQQISHLVIMHEEG UniprotNo.: EVDGKAIPDLTAPVAAVQAAVSNLVRVGKE P18206 TVQTTEDQILKRDMPPAFIKVENACTKLVQ Length: AAQMLQSDPYSVPARDYLIDGSRGILSGTS a)isoform1: DLLLTFDEAEVRKIIRVCKGILEYLTVAEV 1,066aa VETMEDLVTYTKNLGPGMTKMAKMIDERQQ b)isoform2: ELTHQEHRVMLVNSMNTVKELLPVLISAMK 1,134aa IFVTTKNSKNQGIEEALKNRNFTVEKMSAE c)isoform3: INEIIRVLQLTSWDEDAWASKDTEAMKRAL 222aa ASIDSKLNQAKGWLRDPSASPGDAGEQAIR QILDEAGKVGELCAGKERREILGTCKMLGQ MTDQVADLRARGQGSSPVAMQKAQQVSQGL DVLTAKVENAARKLEAMTNSKQSIAKKIDA AQNWLADPNGGPEGEEQIRGALAEARKIAE LCDDPKERDDILRSLGEISALTSKLADLRR QGKGDSPEARALAKQVATALQNLQTKTNRA VANSRPAKAAVHLEGKIEQAQRWIDNPTVD DRGVGQAAIRGLVAEGHRLANVMMGPYRQD LLAKCDRVDQLTAQLADLAARGEGESPQAR ALASQLQDSLKDLKARMQEAMTQEVSDVFS DTTTPIKLLAVAATAPPDAPNREEVFDERA ANFENHSGKLGATAEKAAAVGTANKSTVEG IQASVKTARELTPQVVSAARILLRNPGNQA AYEHFETMKNQWIDNVEKMTGLVDEAIDTK SLLDASEEAIKKDLDKCKVAMANIQPQMLV AGATSIARRANRILLVAKREVENSEDPKFR EAVKAASDELSKTISPMVMDAKAVAGNISD PGLQKSFLDSGYRILGAVAKVREAFQPQEP DFPPPPPDLEQLRLTDELAPPKPPLPEGEV PPPRPPPPEEKDEEFPEQKAGEVINQPMMM AARQLHDEARKWSSKGNDIIAAAKRMALLM AEMSRLVRGGSGTKRALIQCAKDIAKASDE VTRLAKEVAKQCTDKRIRTNLLQVCERIPT ISTQLKILSTVKATMLGRTNISDEESEQAT EMLVHNAQNLMQSVKETVREAEAASIKIRT DAGFTLRWVRKTPWYQ (SEQIDNO12) b)Isoform2: MPVFHTRTIESILEPVAQQISHLVIMHEEG EVDGKAIPDLTAPVAAVQAAVSNLVRVGKE TVQTTEDQILKRDMPPAFIKVENACTKLVQ AAQMLQSDPYSVPARDYLIDGSRGILSGTS DLLLTFDEAEVRKIIRVCKGILEYLTVAEV VETMEDLVTYTKNLGPGMTKMAKMIDERQQ ELTHQEHRVMLVNSMNTVKELLPVLISAMK IFVTTKNSKNQGIEEALKNRNFTVEKMSAE INEIIRVLQLTSWDEDAWASKDTEAMKRAL ASIDSKLNQAKGWLRDPSASPGDAGEQAIR QILDEAGKVGELCAGKERREILGTCKMLGQ MTDQVADLRARGQGSSPVAMQKAQQVSQGL DVLTAKVENAARKLEAMTNSKQSIAKKIDA AQNWLADPNGGPEGEEQIRGALAEARKIAE LCDDPKERDDILRSLGEISALTSKLADLRR QGKGDSPEARALAKQVATALQNLQTKTNRA VANSRPAKAAVHLEGKIEQAQRWIDNPTVD DRGVGQAAIRGLVAEGHRLANVMMGPYRQD LLAKCDRVDQLTAQLADLAARGEGESPQAR ALASQLQDSLKDLKARMQEAMTQEVSDVFS DTTTPIKLLAVAATAPPDAPNREEVFDERA ANFENHSGKLGATAEKAAAVGTANKSTVEG IQASVKTARELTPQVVSAARILLRNPGNQA AYEHFETMKNQWIDNVEKMTGLVDEAIDTK SLLDASEEAIKKDLDKCKVAMANIQPQMLV AGATSIARRANRILLVAKREVENSEDPKFR EAVKAASDELSKTISPMVMDAKAVAGNISD PGLQKSFLDSGYRILGAVAKVREAFQPQEP DFPPPPPDLEQLRLTDELAPPKPPLPEGEV PPPRPPPPEEKDEEFPEQKAGEVINQPMMM AARQLHDEARKWSSKPGIPAAEVGIGVVAE ADAADAAGFPVPPDMEDDYEPELLLMPSNQ PVNQPILAAAQSLHREATKWSSKGNDIIAA AKRMALLMAEMSRLVRGGSGTKRALIQCAK DIAKASDEVTRLAKEVAKQCTDKRIRTNLL QVCERIPTISTQLKILSTVKATMLGRTNIS DEESEQATEMLVHNAQNLMQSVKETVREAE AASIKIRTDAGFTLRWVRKTPWYQ (SEQIDNO13) c)Isoform3: MPPAFIKVENACTKLVQAAQMLQSDPYSVP ARDYLIDGSRGILSGTSDLLLTFDEAEVRK IIRVCKGILEYLTVAEVVETMEDLVTYTKN LGPGMTKMAKMIDERQQELTHQEHRVMLVN SMNTVKELLPVLISAMKIFVTTKNSKNQGI EEALKNRNFTVEKMSAEINEIIRVLQLTSW DEDAWASKVRVLSGEISKIPNSPWLGVLIG TCLILYLVIFVA (SEQIDNO14) Pre-pro MDYLLMIFSLLFVACQGAPETAVLGAELSA Endothelin1 VGENGGEKPTPSPPWRLRRSKRCSCSSLMD (ET-1) KECVYFCHLDIIWVNTPEHVVPYGLGSPRS UniprotNo.: KRALENLLPTKATDRENRCQCASQKDKKCW P05305 NFCQAGKELRAEDIMEKDWNNHKKGKDCSK Length: LGKKCIYQQLVRGRKIRRSSEEHLRQTRSE 212aa TMRNSVKSSFHDPKLKGKPSRERYVTHNRA Lengthof HW fragments (SEQIDNO15) ofpre-pro a)aminoacidsequenceof Endothelin: pro-ET-1: a)202aa APETAVLGAELSAVGENGGEKPTPSPPWRL b)21aa RRSKRCSCSSLMDKECVYFC c)45aa HLDIIWVNTPEHWPYGLGSPRSKRALENLL d)38aa PTKATDRENRCQCASQKDKKCWNFCQAGKE LRAEDIMEKDWNNHKKGKDCSKLGKKCIYQ QLVRGRKIRRSSEEHLRQTRSETMRNSVKS SFHDPKLKGKPSRERYVTHNRAHW (SEQIDNO16) b)aminoacidsequenceofET-1: CSCSSLMDKECVYFCHLDIIW (SEQIDNO17) c)aminoacidsequenceof CT-ET-1: RSSEEHLRQTRSETMRNSVKSSFHDPKLKG KPSRERYVTHNRAHW (SEQIDNO18) d)aminoacidsequenceof Big-ET-1: CSCSSLMDKECVYFCHLDIIWVNTPEHWPY GLGSPRS (SEQIDNO19) Procalcitonin APFRSALESSPADPATLSEDEARLLLAALV (PCT) QDYVQMKASELEQEQEREGSSLDSPRSKRC length116aa GNLSTCMLGTYTQDFNKFHTFPQTAIGVGA PGKKRDMSSDLERDHRPHVSMPQNAN (SEQIDNO20) SRMpeptide LLGIETPLPK ofICAM1 (SEQIDNO21) Length: 10aa SRMpeptide FSVVYAK ofAHSG (SEQIDNO22) Length: 7aa SRMpeptide VQNECPGITR ofCPN1 (SEQIDNO23) Length: 10aa SRMpeptide AIGLPEELIQK ofFABP1 (SEQIDNO24) Length: 11aa SRMpeptide DGYLFQLLR ofHRG (SEQIDNO25) Length: 9aa SRMpeptide CGLGINSR ofPIGR (SEQIDNO26) Length: 8aa SRMpeptide EQGQNLAR ofRAP1A/ (SEQIDNO27) RAP1B/RA PBL Length: 8aa SRMpeptide FVFGTTPEDILR ofTHBS1 (SEQIDNO28) Length: 12aa SRMpeptide GNDIIAAAK ofVCL (SEQIDNO29) Length: 9aa