Molecular markers for diagnosing active tuberculous diseases and application thereof

A molecular marker, tuberculosis-like technology, applied in the field of molecular biology, can solve the problems of less tuberculosis reports, no B12 and tuberculosis, etc.

Active Publication Date: 2017-12-08
GUANGDONG MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a large number of studies have confirmed that CD272 is involved in the occurrence and development of different infectious diseases, but there are few reports on its function with tuberculosis
There is no research report on the relationship between B12 and tuberculosis

Method used

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  • Molecular markers for diagnosing active tuberculous diseases and application thereof
  • Molecular markers for diagnosing active tuberculous diseases and application thereof
  • Molecular markers for diagnosing active tuberculous diseases and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Expression of embodiment 1, miR-16, B12 mRNA and CD272 mRNA in active pulmonary tuberculosis patient and healthy volunteer PBMC

[0024] 47 cases of active pulmonary tuberculosis patients diagnosed by "WS 288-2008 Tuberculosis Diagnostic Criteria", including 15 cases of sputum smear positive, 32 cases of sputum smear negative, 52 cases of healthy volunteers (HV), meet the clinical symptoms of tuberculosis and chest imaging There was no obvious abnormality in the examination, and there was no significant difference in gender and age between the tuberculosis patient group and the tuberculosis patient group.

[0025] 1. PBMC separation

[0026] Collect peripheral venous blood from TB patients and HV healthy subjects, add lymphocyte separation medium with the same volume as the blood sample; centrifuge horizontally at 1200rpm for 20min at room temperature; carefully draw the second layer of white lymphocyte layer into another sterile centrifuge tube, and use Resuspend and ...

Embodiment 2

[0039] Example 2, miR-16, the relative expression of B12 mRNA and CD272 mRNA is related to the classification of active pulmonary tuberculosis

[0040] The classification criteria for pulmonary tuberculosis patients are: sputum smear microscope examination positive for tuberculosis (sputum smear positive, SS+): tuberculosis acid-fast bacilli can be seen under the sputum smear microscope or a sputum culture test is positive for tuberculosis; sputum smear microscopy Negative tuberculosis test (sputum smear negative, SS-): at least 3 sputum smears and one sputum culture test were negative for tuberculosis.

[0041] Combined with the sputum examination of pulmonary tuberculosis patients, 15 cases were positive in sputum smear and 32 cases were negative in sputum smear, the correlation between the relative expression of miR-16, B12mRNA and CD272 mRNA and the classification of pulmonary tuberculosis was analyzed. The results are as follows: figure 2 As shown, in patients with activ...

Embodiment 3

[0042] Example 3. Auxiliary diagnosis of active pulmonary tuberculosis by combination of miR-16, B12 and CD272 mRNA

[0043] According to the above test results, 52 peripheral blood samples of active pulmonary tuberculosis and 50 peripheral blood samples of normal healthy volunteers were taken, and Real-time PCR was used to detect the single index and the combination of three indexes in PBMC. The relative expression level of miR-16 in sample PBMC ≥ 1.3, the relative expression level of B12 mRNA ≥ 0.5, and the relative expression level of CD272 mRNA ≤ 20 are the criteria for confirming active pulmonary tuberculosis patients; Tuberculosis Diagnostic Criteria "diagnosed as the gold standard, to judge the accuracy of index detection, accuracy = the correct number of index detection samples / total number of detections.

[0044] Table 1. Accuracy results of single index diagnosis and combined diagnosis of tuberculosis patients and healthy volunteers

[0045]

[0046] The result...

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Abstract

The invention discloses molecular makers for diagnosing active tuberculous diseases and applications thereof. The invention proves that miR-16, B12 and CD272 can be used as the molecular markers for diagnosing the active tuberculous diseases, diagnosis with a combination of the three can improve the diagnosis accuracy rate (more than 90%) of the tuberculous diseases, and the diagnosis accuracy rate is far superior to that of diagnosis with a single marker, so that the molecular markers are suitable for screening, diagnosis and efficacy evaluation for the active tuberculous diseases.

Description

technical field [0001] The invention belongs to the field of molecular biology, and more specifically relates to a molecular marker for diagnosing active pulmonary tuberculosis and its application. Background technique [0002] It is estimated that 1 / 3 of the world's population is infected with Mycobacterium tuberculosis (MTB), and about 5% to 10% of the infected population will eventually develop tuberculosis (TB). my country is one of the 22 countries with a high burden of TB in the world. one. According to the 2014 WHO global TB control report, there are about 9 million new TB patients each year in the world, and about 1.5 million of them die. In my country, there are more than 1 million new TB patients every year. The TB epidemic presents a high infection rate, high prevalence rate, and high resistance. There are large differences in drug rate and regional prevalence rate, rural epidemic situation is higher than that in urban areas, middle-aged and elderly people are more...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178
Inventor 曾今诚袁耀钦林东子林碧华周克元张海涛刘新光黄明元刘乾坤梅月志
Owner GUANGDONG MEDICAL UNIV
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