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Application of mycobacterium tuberculosis protein in preparation of product for diagnosing latent tuberculosis infectors and/or active pulmonary tuberculosis

An active pulmonary tuberculosis and protein technology, applied in the field of biomedicine, can solve the problems of inability to differentially diagnose latent Mycobacterium tuberculosis and low positive detection rate, and achieve the effect of good specificity and high sensitivity

Active Publication Date: 2020-08-11
BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is aimed at the low positive detection rate of products for differential diagnosis of latent tuberculosis infection and / or active pulmonary tuberculosis in the prior art, and the problem that latent Mycobacterium tuberculosis infection cannot be differentially diagnosed. Use of a Mycobacterium tuberculosis protein in preparing products for diagnosing tuberculosis latent infection and / or active pulmonary tuberculosis

Method used

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  • Application of mycobacterium tuberculosis protein in preparation of product for diagnosing latent tuberculosis infectors and/or active pulmonary tuberculosis
  • Application of mycobacterium tuberculosis protein in preparation of product for diagnosing latent tuberculosis infectors and/or active pulmonary tuberculosis

Examples

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Effect test

Embodiment 1

[0037] Example 1: Collection and processing of samples

[0038] The sample detected by the present invention is serum, and the collection method is to collect the blood of the subject on an empty stomach, use an ordinary biochemical test tube without anticoagulant, let it stand at room temperature for natural coagulation, centrifuge at 3000 rpm for 10 minutes at room temperature, and collect the serum fraction. Packed in cryopreservation tubes and stored in a -80°C low-temperature refrigerator for use in experiments.

[0039] The samples used in the present invention are divided into two groups: a latent infection group and an active tuberculosis patient group; wherein, the latent infection group refers to those who have no clinical complaint symptoms, TST test and T-SPOT test are positive, and chest X-ray films are negative; The active tuberculosis patient group refers to the confirmed tuberculosis patients with positive smear or culture, who have not started or just started ...

Embodiment 2

[0041] Embodiment 2: Utilize ELISA experiment to the detection of sample

[0042] 1. Establish various buffers, diluents, reaction solutions and stop solutions required for ELSIA experiments:

[0043] (1) Coating buffer (pH9.6, 0.05mol / L carbonate buffer):

[0044] Na 2 CO 3 1.59g, NaHCO 3 Add double distilled water to 2.93g to 1000ml, adjust pH to 9.6;

[0045] (2) PBS solution with pH7.4:

[0046] NaCl 8.0g, KCl 0.2g, KH 2 PO 4 0.2g, Na 2 HPO 4 12H 2 O 2.9g,

[0047] Add double distilled water to 1000ml, adjust pH to 7.4;

[0048] (3) PBST washing solution with pH7.4:

[0049] Add 0.5ml Tween-20 to 1000ml PBS solution, adjust the pH to 7.4;

[0050] (4) Blocking solution (pBS solution containing skimmed milk powder at pH 7.4):

[0051] Add 5g skimmed milk powder to 100ml PBS solution;

[0052] (5) Substrate buffer: 0.2M NaH 2 PO 4 (28.4g / L) 25.7ml,

[0053] 0.1M citric acid (19.2g / L) 24.3ml, add 50ml of distilled water.

[0054] (6) TMB (tetramethylbenz...

Embodiment 3

[0076] Example 3: Specificity and sensitivity of the method for differential diagnosis of active tuberculosis patients and latent infections using ELISA

[0077] Adopt clinical diagnosis as 92 parts of serum of patients with active pulmonary tuberculosis, 93 parts of serum of latent infection; Utilize the ELISA method among the embodiment 1 and 2 to detect, judge whether the test subject is active pulmonary tuberculosis positive by the judgment standard of embodiment 2 , the result is as figure 2 As shown, the specificity of the result analysis was 83.87%, and the sensitivity was 79.35%. figure 2 The abscissa 1-specificity represents the false positive rate, and the ordinate sensitivity represents the true positive rate.

[0078] The above results prove that the present invention detects the serum antibody response levels of three different Mycobacterium tuberculosis proteins, can better differentiate and diagnose latent tuberculosis infection patients and active tuberculos...

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Abstract

The invention provides application of mycobacterium tuberculosis protein in preparation of products for diagnosing latent tuberculosis infectors and active pulmonary tuberculosis. The method providedby the invention can be used for detecting the levels of three different mycobacterium tuberculosis protein serum antibody responses at the same time, so that latent tuberculosis infectors and activetuberculosis patients can be better identified and diagnosed, the sensitivity is higher, and the specificity is better.

Description

[0001] This application is a divisional application of an invention patent application with an application date of December 30, 2016 and an application number of 2016112595642. technical field [0002] The invention relates to the field of biomedicine, in particular to the use of mycobacterium tuberculosis protein in preparing products for diagnosing tuberculosis latent infection and / or active pulmonary tuberculosis. Background technique [0003] Tuberculosis caused by Mycobacterium tuberculosis is still one of the diseases that seriously threaten human health. According to the latest report of the World Health Organization, there were 10.4 million new cases of tuberculosis in the world in 2015, and about 1.8 million people died of tuberculosis. The number of people is 918,000, the death toll is 38,000, and the total number of cases ranks fourth in the world. Whether in the world or in China, there is still a long way to go in the prevention and control of tuberculosis, espe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/6854G01N33/5695G01N2469/20Y02A50/30
Inventor 李传友曹树辉陈艳清
Owner BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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