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Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of latent tuberculosis infection

A pulmonary tuberculosis and latent technology, applied in the field of biomedicine, can solve the problems that latent tuberculosis infection cannot be diagnosed and the positive detection rate is low.

Active Publication Date: 2014-04-02
TB HEALTHCARE BIOTECHNOLOGY (GUANGDONG) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antigens currently targeted for serological diagnosis of Mycobacterium tuberculosis, such as antigen 5, 38KD antigen, 30 / 31KD antigen, and antigen 60, all have problems such as low positive detection rate and cross-immunity with other mycobacteria. It has certain value in monitoring, prompting recurrence, judging prognosis and general screening of high-risk groups, but it still cannot be used for the diagnosis of latent tuberculosis infection

Method used

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  • Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of latent tuberculosis infection
  • Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of latent tuberculosis infection
  • Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of latent tuberculosis infection

Examples

Experimental program
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Effect test

Embodiment 1

[0072] Embodiment 1, the preparation of protein chip

[0073] (1) Preparation of Mycobacterium tuberculosis antigenic proteins Rv2693c and Rv1984c

[0074] 1. Express

[0075] The genetically engineered Saccharomyces cerevisiae uses galactose to induce overexpression, and the specific process is as follows:

[0076] First, from Mycobacterium M. tuberculosis H37Rv strain (Beijing strain), the gene coding fragments of protein Rv2693c and Rv1984c were de novo cloned by PCR, and the fragments were connected to the pDONR221 vector ( purchased from Invitrogen), transformed into Escherichia coli DH5-Alpha and amplified, the extracted vector was then replaced by LR enzyme (Invitrogen) to the modified pEGH-A vector capable of expressing GST tags (the vector is in "Jian Zhu, Heng Zhu , et al: J.Virol.May2009vol.83no.105219-5231", the public can obtain from Guangdong Tibikang Biotechnology Co., Ltd.), and then transformed into E. coli DH5-Alpha to amplify, and extract the plasmid for t...

Embodiment 2

[0135] Example 2. Application of protein chip for auxiliary diagnosis of latent pulmonary tuberculosis infection

[0136] (1) Preparation of serum samples to be tested

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Abstract

The invention provides mycobacterium tuberculosis protein Rv2693c and / or Rv1984c in development and / or designing of products capable of discrimination, diagnosis, auxiliary diagnosis, screening and / or auxiliary screening of latent tuberculosis infection. The invention further provides protein chips prepared from the two mycobacterium tuberculosis proteins. The protein chips prepared in the invention are used for detecting the levels of IgM antibodies respectively corresponding to the twp proteins in serum of a patient with latent tuberculosis infection and of a normal person, and detection results of the antibodies respectively corresponding to the three protein are analyzed together so as to determine whether an examined person suffers from latent tuberculosis infection; detection results show that optimal operating points of the protein chips provided by the invention in auxiliary diagnosis of latent tuberculosis infection have specificity of 80.3% and sensitivity of 75.6%, both higher than indexes of diagnosis of latent tuberculosis infection in the prior art.

Description

technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to the use of mycobacterium tuberculosis protein in preparing products for diagnosing and / or assisting in diagnosing diseases. Background technique [0002] For centuries, tuberculosis has continued to be a global public health problem that cannot be ignored. At present, one-third of the world's population is carrying Mycobacterium tuberculosis. In 2010 alone, there were 8.8 million new tuberculosis cases and 1.45 million deaths. On average, one person died of tuberculosis in less than 22 seconds. Tuberculosis is the highest infectious disease The number one death toll. my country is one of the 22 countries with a high burden of tuberculosis in the world. The number of tuberculosis patients ranks second in the world, and the number of infected people exceeds 500 million. In 2010 alone, there were 900,000 to 1.2 million new cases, accounting for the total number of new cases in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569C07K14/35
CPCG01N33/5695G01N33/6884G01N33/6893G01N2800/12
Inventor 陶生策张先恩毕利军邓教宇张治平江何伟周盈谷晶郭书娟张鸿泰王绪德王靖方侯剑周杰杨姗姗赵斐
Owner TB HEALTHCARE BIOTECHNOLOGY (GUANGDONG) CO LTD
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