BIOMARKERS FOR THE DIAGNOSIS OF INVASIVE FUNGAL INFECTIONS
20210349089 · 2021-11-11
Assignee
Inventors
- Janin Schulte (Hennigsdorf, DE)
- Anne Incamps (Saint Victor la Coste, FR)
- Markus Alexander Weigand (Wettenberg, DE)
- Thorsten Brenner (Mörlenbach, DE)
Cpc classification
G01N33/53
PHYSICS
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 the diagnosis, prognosis, risk assessment and/or therapy monitoring of a fungal infection in a subject, comprising the step of determining the level of at least one biomarker selected from the group consisting of intercellular adhesion molecule 1 (ICAM1), 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) and endothelin 1 (ET-1) in a sample of said subject, wherein said level of the at least one biomarker is indicative for the presence, the risk of getting, the severity and/or the type of fungal infection in said subject.
2. The method of claim 1, wherein the method is for the diagnosis of an invasive fungal infection in a subject, comprising the step of determining the level of at least one biomarker selected from the group consisting of ICAM1, AHSG, CPN1, FABP1, HRG, PIGR, RAP1, THBS1, VCL and ET-1 in said sample of said subject, wherein said level of the at least one biomarker is indicative for the presence of an invasive fungal infection in said subject.
3. The method of claim 1, wherein the method is for risk assessment and/or therapy monitoring, wherein the method comprises the step of determining the level of at least one biomarker selected from the group consisting of ICAM1, AHSG, CPN1, FABP1, HRG, PIGR, RAP1, THBS1, VCL and ET-1 or fragments thereof in said sample of said subject, wherein the level of the at least one biomarker is indicative of a need of said subject to receive anti-fungal treatment.
4. The method of claim 1, wherein the level of at least the biomarkers a) ICAM1 and THBS1; b) ICAM1 and VCL; or c) ICAM1, THBS1 and VCL are determined.
5. The method of claim 2, wherein the invasive fungal infection is an acute, serious fungal infection, and wherein the acute, serious fungal infection is a systemic fungal infection, a fungemia or a multifocal infection.
6. The method of claim 1, wherein the level of said biomarker(s) is determined after the subject is diagnosed of having or getting a fungal infection or the subject is diagnosed to be in a critical disease state and/or after admission of the subject to a medical site.
7. The method of claim 1 wherein said level of the at least one biomarker is compared to a reference value of said at least one biomarker, wherein (i) when the biomarker is selected from the group consisting of ICAM1, FABP1, PIGR and ET-1, a level above said reference value in the sample of said subject is indicative for the presence of an invasive fungal infection in the subject; or (ii) when the biomarker is selected from the group consisting of AHSG, CPN1, HRG, RAP1, THBS1 and VCL, a level below said reference value in the sample of said subject is indicative for the presence of an invasive fungal infection in the subject.
8. The method of claim 7, wherein said reference value is the level of the respective biomarker in one or more samples of a reference subject or a population of reference subjects without said invasive fungal infection and without a fungal colonization.
9. The method of claim 7, wherein said reference value is derived from the level of the respective biomarker in (a) sample(s) of a reference subject or a population of reference subjects without said invasive fungal infection and which has/have a fungal colonization.
10. The method of claim 1, wherein the subject is a subject having an increased risk of getting or having an invasive fungal infection.
11. The method of claim 10, wherein said subject is in a critical disease state.
12. The method of claim 1, wherein said subject is a subject after organ transplantation such as liver transplantation.
13. The method of claim 11, wherein said subject is a subject selected from the group consisting of (i) a patient having at least one chronic or acute viral or bacterial infection; (ii) a patient having a mixed bacterial and viral infection; (iii) a patient having an immune suppression, impaired immune response or dysregulated immune system.
14. The method of claim 1, wherein additionally the level of one or more further biomarker and/or clinical score(s) and/or clinical parameter and/or infection parameter is determined
15. The method of claim 14, wherein the biomarkers ICAM1 and PCT are determined.
16. The method of claim 1, wherein the level of the biomarker is determined by mass spectrometry or in an immunoassay.
17. The method of claim 1, further comprising the diagnosis and/or risk stratification for the course and/or the severity of a fungal infection, in the sample of the patient as an accompaniment to a therapy; wherein said therapy is adjusted comprising administration of appropriate anti-infectious therapeutic agents.
18. An antifungal agent for use in treating an invasive fungal infection in a subject, wherein said antifungal agent is administered to said subject if an invasive fungal infection has been diagnosed or predicted in said subject by the method of claim 1.
Description
DESCRIPTION OF THE DRAWINGS
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[0419] 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
[0420] 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).
[0421] 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).
[0422] Immunoassays. Plasma concentrations of β-D-glucan (BD) were measured using the Glucatell®-Kit (Pyroquant Diagnostik GmbH) according to the manufacturer's instructions. In all 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
[0423] 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
[0424] 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.—
[0425] Clinical Microbiology.
[0426] Blood Culture:
[0427] 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, Wis., USA) and automated antimicrobial susceptibility testing (VITEK 2).
[0428] Culture-Based Diagnostic Procedures in Tracheal Secretion, Wound Swabs and Drainage Fluids:
[0429] 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 48 h, except the Schaedler-KV bi-plates, which were incubated at 37° C. in an anaerobic chamber (GasPak; Becton, Dickinson, Franklin Lakes, N.J.) for 48 h 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
[0430] 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.
[0431] 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).
[0432] Statistical Analyses.
[0433] 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: ***.
[0434] Multiple comparison analysis has been performed by one-way analysis of variance (ANOVA) followed by a Dunnett's post hoc test.
[0435] Results
[0436] 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.
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*
[0437] Data are presented as either number (with the corresponding percentage value) or median (with accompanying quartiles).
[0438] 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
[0439] Fungal Pathogens and Infection Sites.
[0440] 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.
[0441] 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.
[0442] 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.
[0443] (1,3)-β-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.
[0444] 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.
[0445] 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.
[0446] 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.
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)
[0447] 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).
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
[0448] Table 3 presents the diagnostic value of ICAM1 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.
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
[0449] 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.
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
[0450] 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.
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
[0451] 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.
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
[0452] 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.
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)
[0453] Table 8 shows the results of the biomarkers PIGR, ICAM1, CPN1, HRG, THBS1, RAP1A, AHSG, VOL and POT, 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.
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
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
[0454] 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).
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
[0455] 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).
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
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
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
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
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
[0456] 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.
[0457]
[0458] 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.
[0459]
[0460]
[0461]
[0462]
[0463]
[0464]
[0465] The
[0466] ICAM1 increases on last time point compared to previous time point if patient died before 90 d, (
[0467] 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.
[0468]
[0469] 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.
[0470]
[0471]
[0472] 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.
[0473]
[0474] 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.
[0475]
[0476]
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)
[0477] Table 17 shows the results of the biomarkers PIGR, ICAM1, HRG, THBS1, RAP1A AHSG, VOL FABP1 and POT, 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.
[0478]
[0479]
[0480] 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.
[0481]
[0482] 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.
[0483]
[0484]
[0485]
[0486]
[0487] 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.
[0488]
[0489] Plasma samples were collected at the onset of septic shock (T0), and 1 day (T1) afterwards and CT-proET-1 was measured.
[0490]
[0491] 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 (
[0492]
[0493] 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.
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)
[0494]
[0495] (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: *.
[0496] (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.
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
[0497]
[0498] 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.
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
[0499]
[0500] Receiver operating characteristic (ROC) analysis with POT and CAM 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.
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
[0501]
[0502] 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.
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
[0503]
[0504] Receiver operating characteristic (ROC) analysis with PCT, ICAM-1, ADM and IL117 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.
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
[0505]
[0506] 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.
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
[0507]
[0508] 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.
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
[0509]
[0510] 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.
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
[0511]
[0512] 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.
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
[0513]
[0514] 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.
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
[0515]
[0516] 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.
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
[0517]
[0518] 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.
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
[0519]
[0520] 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.
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
[0521]
[0522] 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.
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
[0523]
[0524] 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.
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
[0525]
[0526] 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.
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
[0527]
[0528] 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.
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
[0529]
[0530] 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.
[0531] Table 36 presents the diagnostic value of the combination of PCT, MR-proADM, sICAM-1 and/or IL-17A 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.
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)
[0532]
[0533] 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.
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)
[0534]
[0535] 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
[0536] 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.
[0537] 2.1 ICAM1 for the detection of an invasive fungal infection following liver transplantation
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) [0538]
[0540] 2.2 ICAM1 for the Detection of Fungal Pathogens in IFI Vs. Fungal Colonization Following Liver Transplantation
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 [0541]
[0542] 2.2 MR-proADM for the Detection of Fungal Pathogens in IFI Vs. Fungal Colonization Following Liver Transplantation
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 [0543]
[0544]
[0545] 2.3 ICAM-1 and MR-proADM for the Detection of an Invasive Fungal Infection Following Liver Transplantation
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 [0546]
TABLE-US-00042 AMINO ACID SEQUENCES Marker Amino acid sequence (SEQ ID NO) Intercellular MAPSSPRPALPALLVLLGALFPGPGNAQTS adhesion VSPSKVILPRGGSVLVTCSTSCDQPKLLGI molecule 1 ETPLPKKELLLPGNNRKVYELSNVQEDSQP (ICAM1) MCYSNCPDGQSTAKTFLTVYWTPERVELAP Uniprot No.: LPSWQPVGKNLTLRCQVEGGAPRANLTVVL P05362 LRGEKELKREPAVGEPAEVTTTVLVRRDHH Length: GANFSCRTELDLRPQGLELFENTSAPYQLQ 532 aa TFVLPATPPQLVSPRVLEVDTQGTVVCSLD GLFPVSEAQVHLALGDQRLNPTVTYGNDSF SAKASVSVTAEDEGTQRLTCAVILGNQSQE TLQTVTIYSFPAPNVILTKPEVSEGTEVTV KCEAHPRAKVTLNGVPAQPLGPRAQLLLKA TPEDNGRSFSCSATLEVAGQLIHKNQTREL RVLYGPRLDERDCPGNWTWPENSQQTPMCQ AWGNPLPELKCLKDGTFPLPIGESVTVTRD LEGTYLCRARSTQGEVTRKVTVNVLSPRYE IVIITVVAAAVIMGTAGLSTYLYNRQRKIK KYRLQQAQKGTPMKPNTQATPP (SEQ ID NO 1) Alpha-2-HS- MKSLVLLLCLAQLWGCHSAPHGPGLIYRQP glycoprotein NCDDPETEEAALVAIDYINQNLPWGYKHTL (AHSG) NQIDEVKVWPQQPSGELFEIEIDTLETTCH Uniprot No.: VLDPTPVARCSVRQLKEHAVEGDCDFQLLK P02765 LDGKFSVVYAKCDSSPDSAEDVRKVCQDCP Length: LLAPLNDTRVVHAAKAALAAFNAQNNGSNF 367 aa QLEEISRAQLVPLPPSTYVEFTVSGTDCVA KEATEAAKCNLLAEKQYGFCKATLSEKLGG AEVAVTCTVFQTQPVTSQPQPEGANEAVPT PVVDPDAPPSPPLGAPGLPPAGSPPDSHVL LAAPPGHQLHRAHYDLRHTFMGVVSLGSPS GEVSHPRKTRTVVQPSVGAAAGPVVPPCPG RIRHFKV (SEQ ID NO 2) Carboxypeptidase MSDLLSVFLHLLLLFKLVAPVTFRHHRYDD N catalytic LVRTLYKVQNECPGITRVYSIGRSVEGRHL chain YVLEFSDHPGIHEPLEPEVKYVGNMHGNEA (CPN1) LGRELMLQLSEFLCEEFRNRNQRIVQLIQD Uniprot No.: TRIHILPSMNPDGYEVAAAQGPNKPGYLVG P15169 RNNANGVDLNRNFPDLNTYIYYNEKYGGPN Length: HHLPLPDNWKSQVEPETRAVIRWMHSFNFV 458 aa LSANLHGGAVVANYPYDKSFEHRVRGVRRT ASTPTPDDKLFQKLAKVYSYAHGWMFQGWN CGDYFPDGITNGASWYSLSKGMQDFNYLHT NCFEITLELSCDKFPPEEELQREWLGNREA LIQFLEQVHQGIKGMVLDENYNNLANAVIS VSGINHDVTSGDHGDYFRLLLPGIYTVSAT APGYDPETVTVTVGPAEPTLVNFHLKRSIP QVSPVRRAPSRRHGVRAKVQPQARKKEMEM RQLQRGPA (SEQ ID NO 3) Fatty acid- MSFSGKYQLQSQENFEAFMKAIGLPEELIQ binding KCKDIKGVSEIVQNGKHFKFTITAGSKVIQ protein NEFTVGEECELETMTGEKVKTVVQLEGDNK (FABP1) LVTTFKNIKSVTELNGDIITNTMTLGDIVF Uniprot No.: KRISKRI P07148 (SEQ ID NO 4) Length: 127 aa Histidine-rich MKALIAALLLITLQYSCAVSPTDCSAVEPE glycoprotein AEKALDLINKRRRDGYLFQLLRIADAHLDR (HRG) VENTTVYYLVLDVQESDCSVLSRKYWNDCE Uniprot No.: PPDSRRPSEIVIGQCKVIATRHSHESQDLR P04196 VIDFNCTTSSVSSALANTKDSPVLIDFFED Length: TERYRKQANKALEKYKEENDDFASFRVDRI 525 aa ERVARVRGGEGTGYFVDFSVRNCPRHHFPR HPNVFGFCRADLFYDVEALDLESPKNLVIN CEVFDPQEHENINGVPPHLGHPFHWGGHER SSTTKPPFKPHGSRDHHHPHKPHEHGPPPP PDERDHSHGPPLPQGPPPLLPMSCSSCQHA TFGTNGAQRHSHNNNSSDLHPHKHHSHEQH PHGHHPHAHHPHEHDTHRQHPHGHHPHGHH PHGHHPHGHHPHGHHPHCHDFQDYGPCDPP PHNQGHCCHGHGPPPGHLRRRGPGKGPRPF HCRQIGSVYRLPPLRKGEVLPLPEANFPSF PLPHHKHPLKPDNQPFPQSVSESCPGKFKS GFPQVSMFFTHTFPK (SEQ ID NO 5) Polymeric MLLFVLTCLLAVFPAISTKSPIFGPEEVNS immunoglobulin VEGNSVSITCYYPPTSVNRHTRKYWCRQGA receptor RGGCITLISSEGYVSSKYAGRANLTNFPEN (PIGR) GTFVVNIAQLSQDDSGRYKCGLGINSRGLS Uniprot No.: FDVSLEVSQGPGLLNDTKVYTVDLGRTVTI P01833 NCPFKTENAQKRKSLYKQIGLYPVLVIDSS Length: GYVNPNYTGRIRLDIQGTGQLLFSVVINQL 764 aa RLSDAGQYLCQAGDDSNSNKKNADLQVLKP EPELVYEDLRGSVTFHCALGPEVANVAKFL CRQSSGENCDVVVNTLGKRAPAFEGRILLN PQDKDGSFSVVITGLRKEDAGRYLCGAHSD GQLQEGSPIQAWQLFVNEESTIPRSPTVVK GVAGGSVAVLCPYNRKESKSIKYWCLWEGA QNGRCPLLVDSEGWVKAQYEGRLSLLEEPG NGTFTVILNQLTSRDAGFYWCLTNGDTLWR TTVEIKIIEGEPNLKVPGNVTAVLGETLKV PCHFPCKFSSYEKYWCKWNNTGCQALPSQD EGPSKAFVNCDENSRLVSLTLNLVTRADEG WYWCGVKQGHFYGETAAVYVAVEERKAAGS RDVSLAKADAAPDEKVLDSGFREIENKAIQ DPRLFAEEKAVADTRDQADGSRASVDSGSS EEQGGSSRALVSTLVPLGLVLAVGAVAVGV ARARHRKNVDRVSIRSYRTDISMSDFENSR EFGANDNMGASSITQETSLGGKEEFVATTE STTETKEPKKAKRSSKEEAEMAYKDFLLQS STVAAEAQDGPQEA (SEQ ID NO 6) Ras-related MREYKLVVLGSGGVGKSALTVQFVQGIFVE protein KYDPTIEDSYRKQVEVDCQQCMLEILDTAG Rap-1A TEQFTAMRDLYMKNGQGFALVYSITAQSTF (RAP1A) NDLQDLREQILRVKDTEDVPMILVGNKCDL Uniprot No.: EDERVVGKEQGQNLARQWCNCAFLESSAKS P62834 KINVNEIFYDLVRQINRKTPVEKKKPKKKS Length: CLLL 184 aa (SEQ ID NO 7) Ras-related MREYKLVVLGSGGVGKSALTVQFVQGIFVE protein KYDPTIEDSYRKQVEVDAQQCMLEILDTAG Rap-1b TEQFTAMRDLYMKNGQGFALVYSITAQSTF (RAP1B) NDLQDLREQILRVKDTDDVPMILVGNKCDL Uniprot No.: EDERVVGKEQGQNLARQWNNCAFLESSAKS P61224 KINVNEIFYDLVRQINRKTPVPGKARKKSS isoform 1 CQLL(SEQ ID NO 8) Length: 184 aa Ras-related MREYKLVVLGSRGVGKSALTVQFVQGIFVE protein KYDPTIEDSYREQVEVDAQQCMLEILDTAG Rap-1b- TEQFTAMRDLYMKNGQGFALVYSITAQSTF like protein NDLQDLREQILRVKDTDDVPMILVGNKCDL (RP1BL) EDERVVGKEQGQNLARQWNNCAFLESSAKS Uniprot No.: KINVNEIFYDLVRQINRKTPVPGKARKKSS A6NIZ1 CQLL Length: (SEQ ID NO 9) 184 aa Thrombospondin-1 a) Isoform 1: (THBS1) MGLAWGLGVLFLMHVCGTNRIPESGGDNSV Uniprot No.: FDIFELTGAARKGSGRRLVKGPDPSSPAFR P07996 IEDANLIPPVPDDKFQDLVDAVRAEKGFLL Length: LASLRQMKKTRGTLLALERKDHSGQVFSVV a)isoform 1: SNGKAGTLDLSLTVQGKQHVVSVEEALLAT 1,170 aa GQWKSITLFVQEDRAQLYIDCEKMENAELD b) isoform 2: VPIQSVFTRDLASIARLRIAKGGVNDNFQG 1,085 aa 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 (SEQ ID NO 10) b) Isoform 2: 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 (SEQ ID NO 11) Vinculin a) Isoform 1: (VCL) MPVFHTRTIESILEPVAQQISHLVIMHEEG Uniprot No.: EVDGKAIPDLTAPVAAVQAAVSNLVRVGKE P18206 TVQTTEDQILKRDMPPAFIKVENACTKLVQ Length: AAQMLQSDPYSVPARDYLIDGSRGILSGTS a) isoform 1: DLLLTFDEAEVRKIIRVCKGILEYLTVAEV 1,066 aa VETMEDLVTYTKNLGPGMTKMAKMIDERQQ b) isoform 2: ELTHQEHRVMLVNSMNTVKELLPVLISAMK 1,134 aa IFVTTKNSKNQGIEEALKNRNFTVEKMSAE c) isoform 3: INEIIRVLQLTSWDEDAWASKDTEAMKRAL 222 aa 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 (SEQ ID NO 12) b) Isoform 2: 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 (SEQ ID NO 13) c) Isoform 3: MPPAFIKVENACTKLVQAAQMLQSDPYSVP ARDYLIDGSRGILSGTSDLLLTFDEAEVRK IIRVCKGILEYLTVAEVVETMEDLVTYTKN LGPGMTKMAKMIDERQQELTHQEHRVMLVN SMNTVKELLPVLISAMKIFVTTKNSKNQGI EEALKNRNFTVEKMSAEINEIIRVLQLTSW DEDAWASKVRVLSGEISKIPNSPWLGVLIG TCLILYLVIFVA (SEQ ID NO 14) Pre-pro MDYLLMIFSLLFVACQGAPETAVLGAELSA Endothelin 1 VGENGGEKPTPSPPWRLRRSKRCSCSSLMD (ET-1) KECVYFCHLDIIWVNTPEHVVPYGLGSPRS Uniprot No.: KRALENLLPTKATDRENRCQCASQKDKKCW P05305 NFCQAGKELRAEDIMEKDWNNHKKGKDCSK Length: LGKKCIYQQLVRGRKIRRSSEEHLRQTRSE 212 aa TMRNSVKSSFHDPKLKGKPSRERYVTHNRA Length of HW fragments (SEQ ID NO 15) of pre-pro a) amino acid sequence of Endothelin: pro-ET-1: a) 202 aa APETAVLGAELSAVGENGGEKPTPSPPWRL b) 21 aa RRSKRCSCSSLMDKECVYFC c) 45 aa HLDIIWVNTPEHWPYGLGSPRSKRALENLL d) 38 aa PTKATDRENRCQCASQKDKKCWNFCQAGKE LRAEDIMEKDWNNHKKGKDCSKLGKKCIYQ QLVRGRKIRRSSEEHLRQTRSETMRNSVKS SFHDPKLKGKPSRERYVTHNRAHW (SEQ ID NO 16) b) amino acid sequence of ET-1: CSCSSLMDKECVYFCHLDIIW (SEQ ID NO 17) c) amino acid sequence of CT-ET-1: RSSEEHLRQTRSETMRNSVKSSFHDPKLKG KPSRERYVTHNRAHW (SEQ ID NO 18) d) amino acid sequence of Big-ET-1: CSCSSLMDKECVYFCHLDIIWVNTPEHWPY GLGSPRS (SEQ ID NO 19) Procalcitonin APFRSALESSPADPATLSEDEARLLLAALV (PCT) QDYVQMKASELEQEQEREGSSLDSPRSKRC length 116 aa GNLSTCMLGTYTQDFNKFHTFPQTAIGVGA PGKKRDMSSDLERDHRPHVSMPQNAN (SEQ ID NO 20) SRM peptide LLGIETPLPK of ICAM1 (SEQ ID NO 21) Length: 10 aa SRM peptide FSVVYAK of AHSG (SEQ ID NO 22) Length: 7 aa SRM peptide VQNECPGITR of CPN1 (SEQ ID NO 23) Length: 10 aa SRM peptide AIGLPEELIQK of FABP1 (SEQ ID NO 24) Length: 11 aa SRM peptide DGYLFQLLR of HRG (SEQ ID NO 25) Length: 9 aa SRM peptide CGLGINSR of PIGR (SEQ ID NO 26) Length: 8 aa SRM peptide EQGQNLAR of RAP1A/ (SEQ ID NO 27) RAP1B/RA PBL Length: 8 aa SRM peptide FVFGTTPEDILR of THBS1 (SEQ ID NO 28) Length: 12 aa SRM peptide GNDIIAAAK of VCL (SEQ ID NO 29) Length: 9 aa