Hematological Parameter for Viral Infection
20230160804 · 2023-05-25
Inventors
Cpc classification
G16H50/20
PHYSICS
International classification
Abstract
The disclosure relates to hematological parameters of viral infection. More specifically, the present disclosure relates to automated volume biomarkers lymph index and monocyte distribution width (MDW) for early detection of Coronavirus infection. The invention also relates to a method, device and computer executable program for early diagnosis of SARS-CoV-2 infection using volume biomarker lymph index and monocyte distribution width (MDW). According to some technical solutions of the present invention, the automated volumetric parameter lymph index and MDW can be used as viral biomarkers to help healthcare workers in the out-patient department or fever clinic to rapidly identify those who might be infected with SARS-CoV-2 and to provide valuable information for triage decision making.
Claims
1. A method for identifying a subject having a viral infection, the method comprising: flowing a body fluid sample obtained from the subject through a flowcell; measuring individual cells of a plurality of cells in the body fluid sample; determining one or more cell population parameter data values in the body fluid sample based on the measuring, wherein the one or more cell population parameter data values comprise a population parameter data value from one or more of lymphocyte, neutrophil or monocyte; and determining whether or not at least one of the one or more cell population parameter data values exceeds a predetermined threshold, wherein if at least one of the one or more cell population parameter data values exceeds a predetermined threshold, the viral infection in the subject is indicated.
2. The method of claim 1, wherein the viral infection is associated with upper respiratory tract illnesses, or wherein the viral infection is a Coronavirus infection.
3. (canceled)
4. The method of claim 2, wherein the Coronavirus is SARS-CoV-2.
5. The method of claim 1, wherein the one or more cell population parameter data values comprises one or more selected from the group consisting of monocyte distribution width (MDW), lymphocyte volume (LV), lymphocyte distribution width (LV-SD), lymphocyte conductivity (LC), lymph index, mean neutrophil volume (MNV), neutrophil distribution width (NDW), or any combinations thereof.
6. The method of claim 5, wherein the one or more cell population parameter data values comprises MDW, and one or more of lymph index or LV-SD, or wherein the cell population parameter data comprises MNV and NDW.
7. (canceled)
8. The method of claim 5, wherein if the values of the MDW and the lymph index are both outside of their respective reference ranges, the viral infection in the subject is indicated.
9. The method of claim 5, wherein if the value of the MDW is greater than 20.27, the viral infection in the subject is indicated, or wherein if the value of the lymph index is greater than 11.3, the viral infection in the subject is indicated.
10-17. (canceled)
18. The method of claim 5, wherein if the value of the LV-SD is greater than 14.41, the viral infection in the subject is indicated.
19. The method of claim 1, wherein the body fluid sample is whole blood.
20. The method of claim 1, wherein the measuring comprises measuring one or more of volume parameter, conductivity parameter, or light scatter parameter, or wherein the measuring comprises measuring light scatter and direct current impedance from individual cells of the plurality of cells.
21. (canceled)
22. The method of claim 5, wherein the lymph index is calculated according to:
lymph index=LV×(LV-SD)/LC.
23. The method of claim 1, wherein the subject is an individual not confirmed of viral infection, wherein the subject is an individual suspected of viral infection, or wherein the subject is negative for viral nuclear acid testing.
24-26. (canceled)
27. A device for identifying a subject having a viral infection, the device comprising: a transducer for measuring cells passing through a flowcell; and one or more processors configured to perform operations comprising: receiving and processing measurement data from the transducer, determining one or more cell population parameter data values based on the measuring, wherein the one or more cell population parameter data values comprise a population parameter data value from one or more of lymphocyte, neutrophil, or monocyte, and determining whether or not at least one of the one or more cell population parameter data values exceeds a predetermined threshold, wherein if at least one of the one or more cell population parameter data values exceeds a predetermined threshold, the viral infection in the subject is indicated.
28. The device of claim 27, wherein the viral infection is associated with upper respiratory tract illnesses, or wherein the viral infection is a Coronavirus infection.
29. (canceled)
30. (canceled)
31. The device of claim 27, wherein the one or more cell population parameter data values comprises one or more selected from the group consisting of monocyte distribution width (MDW), lymphocyte volume (LV), lymphocyte distribution width (LV-SD), lymphocyte conductivity (LC), lymph index, mean neutrophil volume (MNV), neutrophil distribution width (NDW), or any combinations thereof.
32-34. (canceled)
35. The device of claim 31, wherein if the value of the MDW is greater than 20.27, the viral infection in the subject is indicated, wherein if the lymph index is greater than 11.3, the viral infection in the subject is indicated, or wherein if the value of the LV-SD is greater than 14.41, the viral infection in the subject is indicated.
36-44. (canceled)
45. The device of claim 27, wherein the measuring comprises measuring one or more of volume parameter, conductivity parameter, or light scatter parameter, or wherein the measuring comprises measuring light scatter and direct current impedance from the cells.
46-51. (canceled)
52. A computer storage media recording an executable program for identifying a subject having a viral infection, characterized in that when the executable program is executed by a processor, the following steps are performed: measuring cells passing through a flowcell; determining one or more cell population parameter data values in the body fluid sample based on the measuring, wherein the one or more cell population parameter data values comprise a population parameter data value from one or more of lymphocyte, neutrophil, or monocyte; and determining whether or not at least one of the one or more cell population parameter data values exceeds a predetermined threshold, wherein if at least one of the one or more cell population parameter data values exceeds a predetermined threshold, the viral infection in the subject is indicated.
53-55. (canceled)
56. The computer storage media of claim 52, wherein the one or more cell population parameter data values comprises one or more selected from the group consisting of monocyte distribution width (MDW), lymphocyte volume (LV), lymphocyte distribution width (LV-SD), lymphocyte conductivity (LC), lymph index, mean neutrophil volume (MNV), neutrophil distribution width (NDW), or any combinations thereof.
57-59. (canceled)
60. The computer storage media of claim 56, wherein if the value of the MDW is greater than 20.27, the viral infection in the subject is indicated, wherein if the value of the lymph index is greater than 11.3, the viral infection in the subject is indicated, or wherein if the value of the LV-SD is greater than 14.41, the viral infection in the subject is indicated.
61-69. (canceled)
70. The computer storage media of claim 52, wherein the measuring comprises measuring one or more of volume parameter, conductivity parameter, or light scatter parameter, or wherein the measuring comprises measuring light scatter and direct current impedance from the cells.
71-76. (canceled)
Description
BRIEF DESCRIPTION OF DRAWINGS
[0048]
[0049]
DETAILED DESCRIPTION
Examples
Materials and Method
Case Selection and Data Collection
[0050] In this case-control study, clinical information including contact history, initial symptoms, routine hematology analysis, chest CT and RT-PCT analysis was collected from a total 128 hospitalized patients of Chinese ethnicity from Feb. 14 to 29, 2020 in The Wuhan Union Hospital, Wuhan China. Based on the criteria complied with the guideline for the diagnosis and treatment of 2019 novel coronavirus (COVID-19) infected pneumonia (sixth version), there were 96 patients (male:female ratio, 40:56) including 68 confirmed cases (contact history, clinical symptoms, CT image resembling viral infection and positive molecular testing) and 28 suspected cases (contact history, clinical symptoms, CT image resembling viral infection and negative molecular testing on admission). 32 individuals without clinical and radiological evidence of viral infection were used as controls.
Classification Criteria
[0051] Based on the criteria complied with the guideline for the diagnosis and treatment of 2019 novel coronavirus (COVID-19) infected pneumonia (sixth version), the classification indicators are as follows:
[0052] (I) Suspected Cases
[0053] Comprehensive analysis combining the following epidemiological history and clinical manifestations:
[0054] 1. Epidemiology History
[0055] (1) A travel history or residence history in Wuhan city and surrounding areas thereof, or a travel history or residence history in other communities with case reports within 14 days before onset of illness;
[0056] (2) A history of contact with a patient with new Coronavirus infection (positive nucleic acid testing) within 14 days before onset of illness;
[0057] (3) A history of contact with a patient with fever or respiratory symptoms from Wuhan and surrounding areas thereof, or from communities with reported cases within 14 days before onset of illness;
[0058] (4) Clustering occurrence of ill.
[0059] 2. Clinical manifestations
[0060] (1) Fever and/or respiratory symptoms;
[0061] (2) Imaging features of new Coronavirus pneumonia;
[0062] (3) Normal or decreased total number of white blood cells, reduced lymphocyte count at the early stage of onset of illness.
[0063] Comply with both any one of the epidemiological history and any two of the clinical manifestations.
[0064] No clear epidemiological history, but comply with 3 of the clinical manifestations.
[0065] (II) Confirmed Cases
[0066] Suspected cases with one of the following etiological evidence:
[0067] 1. Positive detection of new coronavirus nucleic acid by real-time fluorescent RT-PCR;
[0068] 2. Highly homologous to known new coronaviruses by viral gene sequencing.
Cell Population Data (CPD) Analysis
[0069] All blood samples were analyzed on a UniCel DxH 800 hematology analyzer (Beckman Coulter, Brea, Calif.) with version 2.0 software within 4 h of collection. This instrument measures CBC with differentials as well as cell morphometric parameters including specific cell volume and distribution of cell volumes within a group of cells, such as mean neutrophil volume (MNV), neutrophil distribution width (NDW), mean monocyte volume (MMV), monocyte distribution width (MDW), mean lymphocyte volume (LV) and lymphocyte distribution width (LV-SD). A simplified lymphocyte CPD, the lymph index, was calculated as LV*LV-SD/LC (lymphocyte conductivity). In addition, 5-angle light-scattering parameters were also collected, including median-angle light scatter, upper median-angle light scatter, lower median-angle light scatter, low-angle light scatter, and axial light loss. The light scatter parameters numerically capture the morphological changes reflected by cellular complexity, granularity, and nuclear structure, with the measurement of AL2 reflecting cell size on the basis of absorbed light.
Statistical Analysis
[0070] All the analysis including ROC are performed with SAS software, the version is 9.4. Since there are total 13 variables for 3 groups for analysis. If the variables are normal distribution, we use one-way ANOVA to test the difference among 3 groups. If the variables are non-normal distribution, we use Kruskal-Wallis to test the difference among 3 groups. If the P-value is less than 0.05, we can conclude that there is a significant difference among 3 groups. Then we use Dwass-Steel-Critchlow-Fligner test for two pairwise comparisons of non-normal distribution variables, if the P-value is less than 0.05, we can conclude that there is a significant difference between 2 groups. In the end, we use Tukey's Studentized Range (HSD) test for two pairwise comparisons of normal distribution variables, if the P-value is less than 0.05, we can conclude that there is a significant difference between 2 groups.
Results
Demographic Data
[0071] Epidemiological information, clinical data, laboratory tests, and radiological characteristics were reviewed from electronic medical records. According to guideline for the diagnosis and treatment of 2019 novel coronavirus (COVID-19) infected pneumonia (sixth version), we prospectively collected and analyzed data from 128 patients (mean age 48.9 years, range 16-88 years; male:female ratio 61:67). There were 68 confirmed cases (common type), 28 suspected cases and 32 cases without clinical and radiological evidence of viral infection as controls. Among 68 confirmed cases, the male to female was 32:36; 68% (46/68) had fever and 47% (32/68) had cough. Among 28 suspected cases, the male to female was 8:20; 68% (19/28) had fever and 57% (16/28) had cough. Note: On follow-up of those suspected patients, 89% (25/28) was subsequently confirmed by positive nuclear testing with COVID-19; 11% (3/28) remained suspicious for COVID-19 infection. This shows that the method of the present disclosure can accurately identify SARS-CoV-2 infected persons at an earlier stage.
Comparison of Conventional Hematologic Parameters and CPD
[0072] As shown in Table 1, no statistically significant differences in conventional hematologic parameters in terms of WBC, percent neutrophils, lymphocytes, monocytes and neutrophil/lymphocyte ratio were observed among all three groups. However, the monocyte distribution width (NDW), lymphocyte distribution width (LV-SD) and lymph index (LV*LV-SD/LC) were significantly increased in both suspected and confirmed group compared to those in the controls (
TABLE-US-00002 TABLE 1 CBC & CPD parameter in three group patients Control (n = 32) Suspect (n = 28) Confirm (n = 68) Mean ± SD Mean ± SD Mean ±S D MDW 19.90 ± 1.95 21.71 ± 2.53 * 22.16 ± 2.26 * * MMV 170.91 ± 8.45 174.39 ± 7.96 174.19 ± 9.10 MNV 146.47 ± 6.51 145.29 ± 5.28 143.78 ± 5.98 NDW 18.62 ± 3.24 17.61 ± 2.53 17.16 ± 1.15 LV 87.84 ± 4.23 88.67 ± 3.27 89.04 ± 4.59 LV-SD 14.62 ± 1.66 15.97 ± 1.56 * 16.25 ± 1.71 * * LC 113.13 ± 7.13 110.71 ± 3.17 111.31 ± 3.23 Lymph-Index 11.36 ± 1.25 12.81 ± 1.55 * 13.02 ± 1.68 * * WBC 6.54 ± 2.65 5.96 ± 2.61 5.65 ± 1.84 NE 61.20 ± 11.21 67.77 ± 12.14 61.90 ± 16.21 LY 25.29 ± 9.62 20.66 ± 10.34 24.89 ± 11.61 MO 9.50 ± 2.39 10.16 ± 2.85 11.00 ± 9.73 NLR 3.37 ± 3.26 5.63 ± 6.53 3.71 ± 3.58 * Suspect vs Control, p < 0.05 * * Confirm vs Control, p < 0.05
The Sensitivity and the Specificity of CPD in Diagnosing COVID-19 Infection
[0073] The sensitivity and the specificity of CPD in predicting COVID-19 infection were then calculated at designated cutoff values. As indicated in Table 2, the MDW and lymph index demonstrated the best sensitivity (78.1% and 84.4%, respectively) and specificity (64.2% and 64.2%, respectively) to detect COVID-19 infection compared to other parameters. ROC curves analysis revealed that MDW and lymph index had the largest areas under the curve (AUC) of 0.77 and 0.79, respectively. With combination of MDW and lymph index, the AUC increases to 0.83 (
TABLE-US-00003 TABLE 2 CPD parameter for Predicting 2019-nCoV infection Cutoff Sensitivity, Specificity, AUC Point % % MDW 0.772 20.27 79.41 59.38 MMV 0.607 172.5 65.6 58.2 MNV 0.397 144.5 40.6 51.5 NDW 0.321 17.23 34.4 42.4 Lymph Index 0.79 11.3 85.29 50 NLR 0.516 2.60 59.4 47.8 MDW + Lymph Index 0.831 85.3 53.1 *AUC, area under the receiver operating characteristic curve.
TABLE-US-00004 TABLE 3 Sensitivity and specificity of MDW and lymph index Sensitivity, Specificity, AUC Cutoff Point % % MDW 0.7730 20.27[20.27, 20.29) 79.41 59.38 MDW 0.7730 20.42[20.42, 20.63) 75 62.5 MDW 0.7730 20.73[20.73, 20.85) 69.12 65.63 Lymph Index 0.7860 11.3[11.3, 11.33) 85.29 50 Lymph Index 0.7860 11.63[11.63, 11.72) 79.41 59.38 Lymph Index 0.7860 11.8[11.8, 11.88) 75 65.63 Lymph Index 0.7860 12.04[12.04, 12.07) 69.12 68.75
DISCUSSION
[0074] Circulating monocytes and lymphocytes are among the first to respond to viral infection. Several previous studies have shown that the volumetric parameters of mononuclear cells are significantly increased during various viral infections [5-7]. Therefore, volumetric increases in these immune cells have potential as viral biomarkers in humans. In current study, we have demonstrated, for the first time, lymph index and monocyte distribution width (MDW) were significantly increased in COVID-19 patients. With designated cutoff values for lymph index and MDW, we achieved the sensitivities of 84.4% and 78.1%, respectively in diagnosing COVID-19 infection. In addition, lymph index in combination with MDW demonstrates excellent diagnostic performance (AUC, 0.83), higher than the AUC with MDW alone and the AUC with lymph index alone. This shows that the combination of MDW and lymph index can more truly reflect the possibility of the subject suffering from SARS-CoV-2 infection, compared with MDW or lymph index alone. Furthermore, we show that lymph index and MDW in suspected patients, like confirmed patients were also significantly increased at the time of admission, suggesting similar pathophysiological process. It is noted that, for the 28 suspected patients tested in this example, they all have negative nucleic acid test at the time of admission, however, 25 of them (89%) were subsequently diagnosed with COVID-19 during the course of disease. This illustrates clinical significance that lymph index and MDW can be used as sensitive screening biomarkers to rapidly identify those suspected individuals before nuclear acid confirmation and to develop proper management plan. We did not see any significant changes in neutrophil volumetric parameters, MNV and NDW, at the time of admission. This is consistent with the observations of recent human studies that neutrophils mainly function as first responders during the innate immune response to acute bacterial infection or sepsis [8-10]
[0075] The whole blood cell analysis plays an important role in healthcare decision making from diagnosis through therapy and prognosis. Currently, the automated hematology analyzers are able to provide not only total leukocyte counts, but also leukocyte volumetric parameters. However, changes in leukocyte numerical parameters, such as total leukocyte count, tend to be extremely variable and nonspecific. In the case of COVID-19, total leukocytes or lymphocytes may be normal or mildly decreased, providing no definitive information for differential diagnosis. Therefore, clinical usefulness of volumetric parameters, lymph index and MDW, offers additional practical advantages. These parameters are generated during automated differential analysis without additional specimen requirements. They can be quantitative and are more accurate since significantly more leukocytes are simultaneously evaluated. Furthermore, they offer more robust turn-around-time and are more cost-effective. These volumetric parameters certainly have potential to become useful viral biomarkers to help healthcare providers in the out-patient department or fever clinic to rapidly identify those who might be infected with COVID-19 and to provide valuable information for triage decision making.
[0076] The scope of the invention is not limited by the specific embodiments described herein. In fact, from the foregoing description and drawings, various modifications of the present invention other than those described herein will be apparent to those skilled in the art. Such modifications are intended to fall within the scope of the appended claims. Furthermore, all embodiments described herein are considered to be broadly applicable and can be combined with any and all other consistent embodiments as appropriate. In addition, if prior art knowledge is not expressly incorporated by reference above, it is expressly incorporated herein in its entirety. A number of publications are cited throughout this document, and all disclosures thereof are incorporated by reference in their entirety.
[0077] The disclosures of the following references are incorporated by reference in their entirety.
REFERENCES
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