Application of Rv1860, Rv0173 and/or Rv1812c protein in preparation of products used for diagnosis of active tuberculosis

A technology for active pulmonary tuberculosis and rv1812c, which is applied in the field of Mycobacterium tuberculosis protein in the preparation of products for diagnosis and/or auxiliary diagnosis of diseases, and can solve the problems of inability to diagnose active pulmonary tuberculosis and low positive detection rate.

Active Publication Date: 2014-04-02
广州博翀生物科技有限公司
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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 active pulmonary tuberculosis

Method used

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  • Application of Rv1860, Rv0173 and/or Rv1812c protein in preparation of products used for diagnosis of active tuberculosis
  • Application of Rv1860, Rv0173 and/or Rv1812c protein in preparation of products used for diagnosis of active tuberculosis
  • Application of Rv1860, Rv0173 and/or Rv1812c protein in preparation of products used for diagnosis of active tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Embodiment 1, the preparation of protein chip

[0083] (1) Preparation of Mycobacterium tuberculosis antigenic proteins Rv1860, Rv0173 and Rv1812c

[0084] 1. Express

[0085] The genetically engineered Saccharomyces cerevisiae uses galactose to induce overexpression, and the specific process is as follows:

[0086] Firstly, from Mycobacterium M.tuberculosis H37Rv strain (Beijing strain), the gene coding fragments of proteins Rv1860, Rv0173 and Rv1812c were de novo cloned by PCR, and the fragments were connected to pDONR221 by BP enzyme from Invitrogen The vector (purchased from Invitrogen) was transformed into Escherichia coli DH5-Alpha for amplification, and the vector was extracted and then replaced with the modified pEGH-A vector capable of expressing GST tags through LR enzyme (Invitrogen) (the vector was found 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 tran...

Embodiment 2

[0147] Example 2. Application of protein chip for auxiliary diagnosis of active pulmonary tuberculosis

[0148] (1) Preparation of serum samples to be tested

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Abstract

The invention provides application of Rv1860, Rv0173 and / or Rv1812c protein in development and / or designing of products capable of discrimination, diagnosis, auxiliary diagnosis, screening and / or auxiliary screening of active tuberculosis. The invention further provides protein chips prepared by using Rv1860, Rv0173 and Rv1812c. The protein chips prepared in the invention are used for detecting the levels of IgG antibodies respectively corresponding to three protein antigens in serum of a patient with active tuberculosis 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 active tuberculosis; detection results show that optimal operating points of the protein chips provided by the invention in auxiliary diagnosis of active tuberculosis have specificity of 83.0% and sensitivity of 73.1%, both higher than indexes of diagnosis of active tuberculosis 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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IPC IPC(8): G01N33/68
CPCG01N33/6884G01N33/6893G01N2800/12
Inventor 毕利军张先恩陶生策邓教宇张治平周盈江何伟谷晶张鸿泰郭书娟王绪德侯剑周杰王靖方杨姗姗赵斐
Owner 广州博翀生物科技有限公司
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