Peripheral blood immune cell subgroup capable of assisting in diagnosing severity of tuberculosis

Abstract

The invention provides a peripheral blood immune cell subset capable of assisting in diagnosing the severity of tuberculosis. Targeted differentiation by the biomarkers of the invention, i.e., the ratio of HLA-DRloS100Ahi mononuclear cells in CD14 + cells (HLA-DRloS100Ahi Mono / CD14 +), the ratio of HLA-DRloS100Ahi mononuclear cells to CD45 + cells (HLA-DRloS100Ahi Mono / CD45 +), the ratio of HLA-DRloS100Ahi mononuclear cells to CD3 + cells (HLA-DRloS100Ahi mononuclear cells to CD4 + cells (HLA-DRloS100Ahi Mono / CD4 +), and / or the ratio of HLA-DRloS100Ahi mononuclear cells to CD8 + cells ( The AUC value of the diagnostic marker for diagnosing a patient with severe pulmonary tuberculosis is greater than 0.8, which indicates that the biomarker screened by the invention has higher diagnostic efficiency on pulmonary tuberculosis.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and particularly relates to a peripheral blood immune cell subset that can assist in diagnosing the severity of tuberculosis. Background Art

[0002] Tuberculosis is a chronic infectious disease and a zoonosis caused by the Mycobacterium tuberculosis complex. The main route of transmission is airborne transmission, and it is mainly divided into two categories: respiratory tuberculosis (such as pulmonary tuberculosis and bronchial tuberculosis) and tuberculosis in other body parts (extrapulmonary tuberculosis). Among them, pulmonary tuberculosis accounts for about 80% of the total cases, and extrapulmonary tuberculosis mainly involves tissues such as the intestine, meninges, lymph nodes, bones, joints, kidneys, and skin (Gautam S S, Acharya B. Advances in diagnosis of Tuberculosis: an update into molecular diagnosis of Mycobacterium tuberculosis [J]. Molecular Biology Reports, 2020, 47(5): 4065 - 4075.). From an epidemiological perspective, tuberculosis remains a major global public health problem in the world. According to WHO statistics, there are 10 million new cases of pulmonary tuberculosis globally every year, including 3.2 million female patients and 1 million child patients. Pulmonary tuberculosis can cause 1.5 million deaths every year (Macneil A, Glaziou P, Sismanidis C, et al. Global epidemiology of Tuberculosis and progress toward meeting global targets - worldwide, 2018 [J]. Morbidity and Mortality Weekly Report, 2020, 69(11): 281 - 285.), and tuberculosis mostly occurs in Southeast Asia and Africa. In addition, once the disease progresses to severe pulmonary tuberculosis, it may cause severe damage to lung tissue, such as extensive cavity formation or fibrosis, which will further affect the cardiopulmonary function and prognosis of patients. Therefore, improving the laboratory diagnosis level of severe pulmonary tuberculosis is of great significance for the treatment of severe pulmonary tuberculosis and the control of the tuberculosis epidemic. Although laboratory detection techniques such as sputum acid - fast bacillus smear microscopy (smear method), sputum mycobacterium culture, and GeneXpert MTB / RIF (Xpert method) are widely used in the diagnosis of pulmonary tuberculosis, relatively few studies have been conducted on the value of these techniques in the diagnosis of severe pulmonary tuberculosis. Therefore, seeking a new breakthrough in the diagnosis of severe pulmonary tuberculosis is an important aspect of controlling tuberculosis. Summary of the Invention

[0003] The present invention has screened out a peripheral blood immune cell subset that can assist in diagnosing the severity of tuberculosis, which shows significant changes in the plasma of patients and has strong diagnostic efficacy for diagnosing severe pulmonary tuberculosis. Based on this, the present invention has been completed.

[0004] In a first aspect, the present invention provides a biomarker for diagnosing severe pulmonary tuberculosis, and the biomarker is HLA-DR lo S100A hi The ratio of monocytes to CD14 + cells (HLA-DR lo S100A hi Mono / CD14 + ), HLA-DR lo S100A hi The ratio of monocytes to CD45 + cells (HLA-DR lo S100A hi Mono / CD45 + ), HLA-DR lo S100A hi The ratio of monocytes to CD3 + cells (HLA-DR lo S100A hi Mono / CD3 + ), HLA-DR lo S100A hi The ratio of monocytes to CD4 + cells (HLA-DR lo S100A hi Mono / CD4 + ), and / or HLA-DR lo S100A hi The ratio of monocytes to CD8 + cells (HLA-DR lo S100A hi Mono / CD8 + ), or one or more of the above.

[0005] Furthermore, the biomarker is derived from the peripheral blood of the patient.

[0006] Furthermore, the severity of the patient's disease is judged according to the level of the biomarker. When HLA-DR lo S100A hi

[0007] Mono / CD14 + > 0.059, HLA-DR lo S100A hiMono / CD45 + >0.0325, HLA - DR lo S100A hi

[0008] Mono / CD3 + >0.0633, HLA - DR lo S100A hi Mono / CD4 + >0.0990 or HLA - DR lo S100A hi

[0009] Mono / CD8 + When >0.1463, it is determined that the patient is a severe pulmonary tuberculosis patient.

[0010] Furthermore, the pulmonary tuberculosis is an infection caused by Mycobacterium tuberculosis.

[0011] Furthermore, the pulmonary tuberculosis includes primary pulmonary tuberculosis, secondary pulmonary tuberculosis, hematogenous disseminated pulmonary tuberculosis, tracheobronchial tuberculosis, tuberculous pleurisy, smear - negative pulmonary tuberculosis, etc.

[0012] Furthermore, the Mycobacterium tuberculosis infection includes: primary infection, secondary infection, extrapulmonary infection.

[0013] In a second aspect, the present invention provides an application of a biomarker in the preparation of a reagent for diagnosing severe pulmonary tuberculosis, and the biomarker is HLA - DR lo S100A hi The proportion of monocytes in CD14 + cells (HLA - DR lo S100A hi Mono / CD14 + ), HLA - DR lo S100A hi The ratio of monocytes to CD45 + cells (HLA - DR lo S100A hi Mono / CD45 + ), HLA - DR lo S100A hi The ratio of monocytes to CD3 + cells (HLA - DR lo S100A hi Mono / CD3 + ), HLA - DR lo S100A hi The ratio of monocytes to CD4 +Ratio of cells (HLA-DR lo S100A hi Mono / CD4 + ) and / or HLA-DR lo S100A hi Ratio of monocytes to CD8 + cells (HLA-DR lo S100A hi Mono / CD8 + ) or more of the above, and the reagent is a reagent containing components capable of detecting the number of HLA-DR lo S100A hi Mono in a patient's biological sample and the number of CD14 + , CD45 + , CD3 + , CD4 + and / or CD8 + cells in the patient's biological sample.

[0014] Furthermore, the biological sample is the patient's peripheral blood.

[0015] Furthermore, the severity of the patient's disease is judged according to the level of the biomarker. When HLA-DR lo S100A hi

[0016] Mono / CD14 + > 0.059, HLA-DR lo S100A hi Mono / CD45 + > 0.0325, HLA-DR lo S100A hi

[0017] Mono / CD3 + > 0.0633, HLA-DR lo S100A hi Mono / CD4 + > 0.0990 or HLA-DR lo S100A hi

[0018] Mono / CD8 + > 0.1463, it is determined that the patient is a severe pulmonary tuberculosis patient.

[0019] In a third aspect, the present invention provides a kit for diagnosing severe pulmonary tuberculosis, and the kit contains the reagent as described in the second aspect of the present invention.

[0020] Furthermore, the kit is a flow cytometry detection kit.

[0021] Beneficial effects

[0022] Through the biomarker of the present invention, namely HLA-DR lo S100A hi The proportion of monocytes in CD14 + cells (HLA-DR lo S100A hi Mono / CD14 + )、HLA-DR lo S100A hi The ratio of monocytes to CD45 + cells (HLA-DR lo S100A hi Mono / CD45 + )、HLA-DR lo S100A hi The ratio of monocytes to CD3 + cells (HLA-DR lo S100A hi Mono / CD3 + )、HLA-DR lo S100A hi The ratio of monocytes to CD4 + cells (HLA-DR lo S100A hi Mono / CD4 + ) and / or HLA-DR lo S100A hi The ratio of monocytes to CD8 + cells (HLA-DR lo S100A hi Mono / CD8 + ) can significantly distinguish severe pulmonary tuberculosis patients from non-severe pulmonary tuberculosis patients. The AUC value of the above diagnostic marker for diagnosing severe pulmonary tuberculosis patients > 0.8, indicating that the biomarker screened by the present invention has a high diagnostic efficacy for pulmonary tuberculosis. Brief description of the drawings

[0023] Figure 1 To screen the levels of HLA-DR lo S100A hi Mono / CD14 + in severe and non-severe pulmonary tuberculosis;

[0024] Figure 2 To screen the levels of HLA-DR lo S100Ahi Mono / CD14 + ROC curve analysis for diagnosing severe pulmonary tuberculosis;

[0025] Figure 3 To screen HLA-DR in the cohort lo S100A hi Mono / CD45 + Levels in severe and non-severe pulmonary tuberculosis;

[0026] Figure 4 To screen HLA-DR in the cohort lo S100A hi Mono / CD3 + Levels in severe and non-severe pulmonary tuberculosis;

[0027] Figure 5 To screen HLA-DR in the cohort lo S100A hi Mono / CD4 + Levels in severe and non-severe pulmonary tuberculosis;

[0028] Figure 6 To screen HLA-DR in the cohort lo S100A hi Mono / CD8 + Levels in severe and non-severe pulmonary tuberculosis;

[0029] Figure 7 To screen HLA-DR in the cohort lo S100A hi Mono / CD45 + ROC curve analysis for diagnosing severe pulmonary tuberculosis;

[0030] Figure 8 To screen HLA-DR in the cohort lo S100A hi ROC curve analysis for diagnosing severe pulmonary tuberculosis with Mono / CD3;

[0031] Figure 9 To screen HLA-DR in the cohort lo S100A hi Mono / CD4 + ROC curve analysis for diagnosing severe pulmonary tuberculosis;

[0032] Figure 10 To screen HLA-DR in the cohort lo S100A hi Mono / CD8 + ROC curve analysis for diagnosing severe pulmonary tuberculosis;

[0033] Figure 11 For HLA-DR in the validation set lo S100A hi Mono / CD14 + Levels in severe and non-severe pulmonary tuberculosis;

[0034] Figure 12 For HLA-DR in the validation set lo S100A hi Mono / CD14 + ROC curve analysis for judging severe pulmonary tuberculosis;

[0035] Figure 13 For HLA-DR in the validation set lo S100A hi Mono / CD45 + Levels in severe and non-severe pulmonary tuberculosis;

[0036] Figure 14 For HLA-DR in the validation set lo S100A hi Mono / CD3 + Levels in severe and non-severe pulmonary tuberculosis;

[0037] Figure 15 For HLA-DR in the validation set lo S100A hi Mono / CD4 + Levels in severe and non-severe pulmonary tuberculosis;

[0038] Figure 16 For HLA-DR in the validation set lo S100A hi Mono / CD8 + Levels in severe and non-severe pulmonary tuberculosis;

[0039] Figure 17 For HLA-DR in the validation set lo S100A hi Mono / CD45 + ROC curve analysis for judging severe pulmonary tuberculosis;

[0040] Figure 18 For HLA-DR in the validation set lo S100A hi Mono / CD3 + ROC curve analysis for judging severe pulmonary tuberculosis;

[0041] Figure 19 For HLA-DR in the validation set loS100A hi Mono / CD4 + ROC curve analysis for judging severe pulmonary tuberculosis;

[0042] Figure 20 For HLA-DR in the validation set lo S100A hi Mono / CD8 + ROC curve analysis for judging severe pulmonary tuberculosis. Specific implementation manners

[0043] The following further describes the specific implementation manners of the present invention. It should be noted here that the description of these implementation manners is used to help understand the present invention, but does not constitute a limitation on the present invention. In addition, the technical features involved in the following described implementation manners can be combined with each other as long as they do not conflict with each other.

[0044] The experimental methods in the following examples are all conventional methods unless otherwise specified, and the test materials used in the following examples are all commercially available through conventional channels unless otherwise specified.

[0045] Example 1 HLA-DR lo S100A hi Mono / CD14 + 、HLA-DR lo S100A hi Mono / CD45 + 、HLA-DR lo S100A hi Mono / CD3 + 、HLA-DR lo S100A hi Mono / CD4 + 、HLA-DR lo S100A hi Mono / CD8 + Screening for judging the severity of pulmonary tuberculosis

[0046] 1. Obtaining venous blood samples

[0047] Venous blood samples of the subjects: 15 healthy controls, 60 non-severe pulmonary tuberculosis patients and 18 severe pulmonary tuberculosis patients.

[0048] Healthy controls: aged ≥ 18 years old; no clinical symptoms of tuberculosis; negative IGRA test; no lung lesions found by chest X-ray or CT; no other serious medical history.

[0049] Non-severe pulmonary tuberculosis: Age ≥ 18 years; having clinical symptoms of tuberculosis; pulmonary lesions found by chest X-ray or CT, but the scope of pulmonary lesions does not reach the level of severe pulmonary tuberculosis.

[0050] Severe pulmonary tuberculosis: Age ≥ 18 years, with unilateral dense pulmonary lesions ≥ 1 / 3 of the unilateral lung field, or bilateral lesions involving ≥ 1 / 2 of the whole lung, or the diameter of the cavity > 4 cm, or combined with a large amount of pleural effusion, and at the same time, active pulmonary tuberculosis patients with PO < 80 mmHg under non-oxygen inhalation state or oxygenation index < 300 under oxygen inhalation state.

[0051] Exclusion criteria: Those receiving immunosuppressive drugs or hormone therapy; autoimmune diseases; viral hepatitis; incomplete clinical data, transferred to another hospital midway or self-abandoned; patients with cancer and active infections.

[0052] 2. Detect HLA-DR in blood specimens lo S100A hi Mono, CD14 + cells, CD45 + cells, CD3 + cells, CD4 + cells and CD8 + cell count

[0053] (1) Preservation of samples: Peripheral blood of patients;

[0054] (2) Detect HLA-DR in blood by flow cytometry analysis system lo S100A hi cell counts of Mono, etc.

[0055] 1) Prepare the sample to be tested;

[0056] 2) Lyse red blood cells;

[0057] 3) Add buffer, vortex, centrifuge, and discard the supernatant;

[0058] 4) Add monoclonal fluorescent antibody mixture;

[0059] 5) Incubate;

[0060] 6) Add buffer, vortex, centrifuge, and discard the supernatant;

[0061] 7) Repeat step 6.

[0062] 8) Add buffer, mix well and wait for detection.

[0063] (3) Detect HLA-DR lo S100A hi Mono, CD14 + cells, CD45 +Cells, CD3 + Cells, CD4 + Cells and CD8 + Cell count.

[0064] 3. Calculate HLA-DR lo S100A hi Mono / CD14 + , HLA-DR lo S100A hi Mono / CD45 + , HLA-DR lo S100A hi Mono / CD3 + , HLA-DR lo S100A hi Mono / CD4 + , HLA-DR lo S100A hi Mono / CD8 +

[0065] Calculate HLA-DR respectively according to the following formula lo S100A hi Mono / CD14 + , HLA-DR lo S100A hi Mono / CD45 + , HLA-DR lo S100A hi Mono / CD3 + , HLA-DR lo S100A hi Mono / CD4 + , HLA-DR lo S100A hi Mono / CD8 + :

[0066]

[0067] Among them, HLA-DR lo S100A hi Mono is the number of HLA-DR low S100A high CD14+ cells in the venous blood sample, CD14 is the number of CD14 + cells in the venous blood sample, CD45 is the number of CD45 + cells in the venous blood sample, CD3 is the number of CD3 +The number of cells, where CD4 is the number of CD4 cells in the venous blood sample + The number of cells, where CD8 is the number of CD8 cells in the venous blood sample + The number of cells.

[0068] 4. Experimental Results

[0069] 4.1. HLA-DR lo S100A hi Mono / CD14 + Used to judge the severity of pulmonary tuberculosis

[0070] HLA-DR lo S100A hi Mono / CD14 + The levels in patients with severe pulmonary tuberculosis were significantly higher than those in patients with non-severe pulmonary tuberculosis (see Figure 1 ). According to the ROC curve analysis, when the cut-off value for judging patients with severe pulmonary tuberculosis by HLA-DR lo S100A hi Mono / CD14 was 0.0590, the area under the curve (AUC) was 0.8690 (95% CI: 0.7609 - 0.9771), the sensitivity was 72.22%, and the specificity was 95.00% (see Figure 2 ).

[0071] 4.2. HLA-DR lo S100A hi Mono / CD45 + 、HLA-DR lo S100A hi Mono / CD3 + 、HLA-DR lo S100A hi Mono / CD4 + 、HLA-DR lo S100A hi Mono / CD8 + Used to judge the severity of pulmonary tuberculosis

[0072] HLA-DR lo S100A hi Mono / CD45 + 、HLA-DR lo S100A hi Mono / CD3 + 、HLA-DR lo S100A hi Mono / CD4 + 、HLA-DR loS100A hi Mono / CD8 + The level in patients with severe pulmonary tuberculosis is significantly higher than that in patients with non-severe pulmonary tuberculosis (see Figure 3 , 4, 5, 6).

[0073] According to the ROC curve analysis, when the critical value for judging patients with severe pulmonary tuberculosis is 0.0325 for HLA-DR lo S100A hi Mono / CD4 + 5, the area under the curve (AUC) is 0.9265 (95% CI: 0.8672 - 0.9858), the sensitivity is 94.44%, and the specificity is 85.48% (see Figure 7 ).

[0074] When the critical value for judging patients with severe pulmonary tuberculosis is 0.0633 for HLA-DR lo S100A hi Mono / CD3 + , the area under the curve (AUC) is 0.9292 (95% CI: 0.8715 - 0.9870), the sensitivity is 83.33%, and the specificity is 87.10% (see Figure 8 ).

[0075] When the critical value for judging patients with severe pulmonary tuberculosis is 0.0990 for HLA-DR lo S100A hi Mono / CD + 4, the area under the curve (AUC) is 0.9274 (95% CI: 0.8667 - 0.9882), the sensitivity is 88.89%, and the specificity is 87.10% (see Figure 9 ).

[0076] When the critical value for judging patients with severe pulmonary tuberculosis is 0.1463 for HLA-DR lo S100A hi Mono / CD8 + , the area under the curve (AUC) is 0.9229 (95% CI: 0.8635 - 0.9824), the sensitivity is 88.89%, and the specificity is 87.10% (see Figure 10 ).

[0077] Example 2 HLA-DR lo S100A hi Mono / CD14 + 、HLA-DR lo S100A hi Mono / CD45 + 、HLA-DRlo S100A hi Mono / CD3 + 、HLA-DR lo S100A hi Mono / CD4 + 、HLA-DR lo S100A hi Mono / CD8 + Verification for judging the severity of pulmonary tuberculosis

[0078] 1. Obtaining venous blood samples

[0079] Venous blood samples of subjects: 15 healthy controls, 63 non-severe pulmonary tuberculosis patients and 18 severe pulmonary tuberculosis patients.

[0080] Healthy controls: aged ≥ 18 years; no clinical symptoms of tuberculosis; negative IGRA test; no pulmonary lesions found by chest X-ray or CT; no other serious medical history.

[0081] Non-severe pulmonary tuberculosis: aged ≥ 18 years; having clinical symptoms of tuberculosis; pulmonary lesions found by chest X-ray or CT, but the scope of pulmonary lesions does not reach the level of severe pulmonary tuberculosis.

[0082] Severe pulmonary tuberculosis: aged ≥ 18 years, with unilateral dense lesions ≥ 1 / 3 of the unilateral lung field, or bilateral lesions involving ≥ 1 / 2 of the whole lung, or a cavity diameter > 4 cm, or combined with a large amount of pleural effusion, and at the same time, active pulmonary tuberculosis patients with PO < 80 mmHg without oxygen inhalation or an oxygenation index < 300 with oxygen inhalation.

[0083] Exclusion criteria: those receiving immunosuppressive drugs or hormone therapy; autoimmune diseases; viral hepatitis; incomplete clinical data, transferred to another hospital or gave up halfway; patients with cancer and active infections.

[0084] 2. Detecting HLA-DR in blood specimens lo S100A hi Mono, CD14 + cells, CD45 + cells, CD3 + cells, CD4 + cells and CD8 + Cell count

[0085] (1) Preservation of samples: peripheral blood of patients;

[0086] (2) Detecting HLA-DR in blood by flow cytometry analysis system lo S100A hi The number of Mono

[0087] 1) Prepare the sample to be tested;

[0088] 2) Lyse red blood cells;

[0089] 3) Add buffer, vortex, centrifuge, and discard the supernatant;

[0090] 4) Add the monoclonal fluorescent antibody mixture;

[0091] 5) Incubate;

[0092] 6) Add buffer, vortex, centrifuge, and discard the supernatant;

[0093] 7) Repeat step 6.

[0094] 8) Add buffer, mix well, and wait for detection.

[0095] (3) Detect HLA-DR lo S100A hi Mono, CD14 + cells, CD45 + cells, CD3 + cells, CD4 + cells and CD8 + cell count.

[0096] 3. Calculate HLA-DR lo S100A hi Mono / CD14 + , HLA-DR lo S100A hi Mono / CD45 + , HLA-DR lo S100A hi Mono / CD3 + , HLA-DR lo S100A hi Mono / CD4 + , HLA-DR lo S100A hi Mono / CD8 +

[0097] Calculate HLA-DR respectively according to the following formulas lo S100A hi Mono / CD14 + , HLA-DR lo S100A hi Mono / CD45 + , HLA-DR lo S100A hi Mono / CD3 +, HLA-DR lo S100A hi Mono / CD4 + , HLA-DR lo S100A hi Mono / CD8 + :

[0098]

[0099] Among them, HLA-DR lo S100A hi Mono is the number of HLA-DR low S100A high CD14+ cells in the venous blood sample, and CD14 is the number of CD14 + cells in the venous blood sample, CD45 is the number of CD45 + cells in the venous blood sample, CD3 is the number of CD3 + cells in the venous blood sample, CD4 is the number of CD4 + cells in the venous blood sample, CD8 is the number of CD8 + cells in the venous blood sample.

[0100] 4. Experimental results

[0101] 4.1. HLA-DR lo S100A hi Mono / CD14 + is used to judge the severity of pulmonary tuberculosis

[0102] HLA-DR lo S100A hi Mono / CD14 + The level in patients with severe pulmonary tuberculosis is significantly higher than that in patients with non-severe pulmonary tuberculosis (see Figure 11 ). When the cut-off value for judging patients with severe pulmonary tuberculosis by HLA-DR lo S100A hi Mono / CD14 + is 0.059, the area under the curve (AUC) is 0.8219 (95% CI: 0.6927 - 0.9510), the sensitivity is 66.7%, and the specificity is 90.48% (see Figure 12 ).

[0103] 4.2 HLA-DR lo S100A hi Mono / CD45 + , HLA-DR lo S100A hiMono / CD3 + 、HLA-DR lo S100A hi Mono / CD4 + 、HLA-DR lo S100A hi Mono / CD8 + For judging the severity of pulmonary tuberculosis

[0104] HLA-DR lo S100A hi Mono / CD45 + 、HLA-DR lo S100A hi Mono / CD3 + 、HLA-DR lo S100A hi Mono / CD4 + 、HLA-DR lo S100A hi Mono / CD8 + The levels in patients with severe pulmonary tuberculosis are significantly higher than those in patients with non-severe pulmonary tuberculosis (see Figure 13 , 14, 15, 16).

[0105] According to the ROC curve analysis, when the critical value for judging a patient as having severe pulmonary tuberculosis is 0.0325 for HLA-DR lo S100A hi Mono / CD45 + The area under the curve (AUC) is 0.8297 (95% CI: 0.7114 - 0.9480), the sensitivity is 61.11%, and the specificity is 81.97% (see Figure 17 ).

[0106] When the critical value for judging a patient as having severe pulmonary tuberculosis is 0.0633 for HLA-DR lo S100A hi Mono / CD3 + The area under the curve (AUC) is 0.8342 (95% CI: 0.7150 - 0.9534), the sensitivity is 61.11%, and the specificity is 88.52% (see Figure 18 ).

[0107] When the critical value for judging a patient as having severe pulmonary tuberculosis is 0.0633 for HLA-DR lo S100A hi Mono / CD4 +When the critical value for judging a patient with severe pulmonary tuberculosis is 0.0990, the area under the curve (AUC) is 0.8370 (95% CI: 0.7202 - 0.9538), the sensitivity is 66.67%, and the specificity is 86.89% (see Figure 19 ).

[0108] When HLA - DR lo S100A hi Mono / CD8 + When the critical value for judging a patient with severe pulmonary tuberculosis is 0.1463, the area under the curve (AUC) is 0.8215 (95% CI: 0.6963 - 0.9467), the sensitivity is 61.11%, and the specificity is 90.16% (see Figure 20 ).

[0109] The results verified the diagnostic efficacy of the biomarker composition of the present invention for judging the severity of pulmonary tuberculosis and determined the feasibility of its application in actual clinical diagnosis.

Claims

1. A biomarker for diagnosing severe pulmonary tuberculosis, the marker being HLA-DR lo S100A hi Monocytes account for CD14 + The ratio of cells (HLA-DR lo S100A hi Mono / CD14 + ), HLA-DR lo S100A hi Monocytes and CD45 + The ratio of cells (HLA-DR lo S100A hi Mono / CD45 + ), HLA-DR lo S100A hi Monocytes and CD3 + The ratio of cells (HLA-DR lo S100A hi Mono / CD3 + ), HLA-DR lo S100A hi Monocytes and CD4 + The ratio of cells (HLA-DR lo S100A hi Mono / CD4 + ) and / or HLA-DR lo S100A hi Monocytes and CD8 + The ratio of cells (HLA-DR lo S100A hi Mono / CD8 + ) The marker according to claim 1 , wherein the marker is derived from the peripheral blood of a patient.

3. The marker according to claim 1, wherein the severity of the disease in the patient is judged according to the level of the marker, when HLA-DR lo S100A hi Mono / CD14 + >0.059, HLA-DR lo S100A hi Mono / CD45 + >0.0325, HLA-DR lo S100A hi Mono / CD3 + >0.0633, HLA-DR lo S100A hi Mono / CD4 + >0.0990 or HLA-DR lo S100A hi Mono / CD8 + When >0.1463, the patient is judged to be a severe pulmonary tuberculosis patient.

4. Use of a biomarker in the preparation of a reagent for diagnosing severe pulmonary tuberculosis, wherein the marker is HLA-DR lo S100A hi Monocytes account for CD14 + The proportion of cells (HLA-DR lo S100A hi Mono / CD14 + ), HLA-DR lo S100A hi Monocytes and CD45 + The ratio of cells (HLA-DR lo S100A hi Mono / CD45 + ), HLA-DR lo S100A hi Monocytes and CD3 + The ratio of cells (HLA-DR lo S100A hi Mono / CD3 + ), HLA-DR lo S100A hi Monocytes and CD4 + The ratio of cells (HLA-DR lo S100A hi Mono / CD4 + ) and / or HLA-DR lo S100A hi Monocytes and CD8 + The ratio of cells (HLA-DR lo S100A hi Mono / CD8 + ), wherein the reagent contains a reagent capable of detecting HLA-DR in a patient's biological sample lo S100A hi Mono number and CD14 in patient biological samples + 、CD45 + 、CD3 + 、CD4 + and / or CD8 + The amount of one or more reagents in a cell.

5. The use according to claim 4, wherein the biological sample is peripheral blood of a patient.

6. The use according to claim 4, wherein the severity of the patient's disease is judged according to the level of the marker, when HLA-DR lo S100A hi Mono / CD14 + >0.059, HLA-DR lo S100A hi Mono / CD45 + >0.0325, HLA-DR lo S100A hi Mono / CD3 + >0.0633、SONG-DR lo S100A hi Mono / CD4 + >0.0990 μm HLA-DR lo S100A hi Mono / CD8 + When >0.1463, the patient is judged to be a severe pulmonary tuberculosis patient.

7. A kit for diagnosing severe pulmonary tuberculosis, comprising the reagent according to claim 4. The kit according to claim 7 , which is a flow cytometry kit.

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