Mycobacterium tuberculosis ESAT-6 recombinant dipolymer, preparation method and application thereof

A technology of ESAT-6 and Mycobacterium tuberculosis, which is applied in the field of bioengineering and diagnostic medicine, can solve the problems of lack of specific antigen, inability to clearly distinguish BCG immunity, environmental mycobacterium infection and pathogenic Mycobacterium tuberculosis infection, application limitations, etc. To achieve the effect of simple and easy method
CN101805397AInactive Publication Date: 2010-08-18SHANGHAI PUBLIC HEALTH CLINICAL CENT
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
SHANGHAI PUBLIC HEALTH CLINICAL CENT
Publication Date
2010-08-18
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the field of biological engineering and diagnostic medicine, in particular to a mycobacterium tuberculosis ESAT-6 recombinant dipolymer, a preparation method thereof, an antigenicity analysis and an application thereof in the serodiagnosis of tuberculosis. The invention uses the DNA of a chimeric gene nucleic acid vaccine HG856A plasmid containing 2 * esat-6 copies as the template, obtains 2 * esat-6 gene segments by PCR amplification, and clones, expresses and purifies an ESAT-6 recombinant dipolymer protein in E.coli. The protein is connected with the two ESAT-6 monomers through amino acid Tyr and Val which are coded by the AccI restriction enzyme cutting site, i.e. GTC TAC, carries six* His tags at the N end, and can be purified by nickel-chelate affinity chromatography. Experiments confirm that the purified recombinant dipolymer has favorable reactogenicity and antigenic specificity. The recombinant dipolymer can be used as an antigen for the early diagnosis of the tuberculosis, including the serodiagnosis of ELISA, ELISPOT or the like, or can be used as an antigen for a skin test.
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Description

technical field

[0001] The invention belongs to the fields of bioengineering and diagnostic medicine. In particular, it relates to a recombinant dimer of Mycobacterium tuberculosis ESAT-6 (rdESAT-6) and its preparation method, antigenicity analysis and application in serological diagnosis of tuberculosis. Background technique

[0002] Tuberculosis is a major public infectious disease of worldwide concern. As early as 1993, the World Health Organization (WHO) declared a state of emergency for the tuberculosis epidemic because of the increasing number of tuberculosis cases worldwide. According to statistics, at present, about 1 / 3 of the world's population (1.86 billion) carries Mycobacterium tuberculosis, with about 8 million new cases and 2 million deaths from tuberculosis every year; there are about 6 million active tuberculosis patients in my country, and there are 250,000 people died from tuberculosis. According to the 2005 infectious disease epidemic situation announced...

Claims

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