Method for detecting mycobacterium tuberculosis by using adaptor technique

A Mycobacterium tuberculosis, technical detection technology, applied in the detection of Mycobacterium tuberculosis using aptamer technology, microbial molecular biology detection, can solve the problem that the cure rate is difficult to improve, the cure rate is low, and it is difficult to deal with multidrug resistance clinical Strains and other problems, to achieve the effect of improving the positive detection rate and simplifying the diagnosis

Inactive Publication Date: 2007-05-09
SHANGHAI PULMONARY HOSPITAL
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Problems solved by technology

[0003] The most important problem in the current tuberculosis control is the low rate of patient detection and low cure rate
In terms of diagnosis, the existing inspection methods have certain limitations, and it is difficult to achieve a fast and accurate diagnosis of tuberculosis.
In terms...

Method used

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  • Method for detecting mycobacterium tuberculosis by using adaptor technique
  • Method for detecting mycobacterium tuberculosis by using adaptor technique
  • Method for detecting mycobacterium tuberculosis by using adaptor technique

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Embodiment 1

[0032] The preparation method of the high-affinity DNA aptamer that can combine with Mycobacterium tuberculosis secretory antigen ESAT-6, CFP-10, MPT53, MPT63, MPT6 protein, take MPT64 as an example, carry out according to the following steps:

[0033] 1. Preparation and purification of MPT64 protein.

[0034]The gene sequence of the MPT64 protein was cloned into a vector according to conventional methods, transfected into E.coli, induced, expressed and purified. The purified protein was analyzed and quantified by SDS-PAGE, and the pure product of the antigen was obtained as the screening target substance.

[0035] The vector involved is pET21a, the C-terminus after expression contains pProEX-HTa, and the N-terminus after expression contains 6×His.

[0036] The vectors involved are techniques known in the art (commercially available).

[0037] The technical measures adopted in the present invention can avoid the low expression level of the protein due to gene construction an...

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Abstract

This invention relates to a method for detecting Mycobacterium tuberculosis by utilizing gamete technique. This invention is aimed to solving the problems of low discovery rate and low curative rate of tuberculosis. The method comprises: utilizing gamete technique, performing bacteria culture or bioengineering method to obtain all kinds of targets of Mycobacterium tuberculosis, obtaining highly compatible specific gametes of the targets by gamete technique screening, and converting the gametes into report gametes for rapid and precise detection of corresponding targets. The method can further combine highly compatible gametes with pathogenetic factors to defunctionalize them, or develop new drugs by finding structural analogues of highly compatible gametes, thus can provides basis for diagnosis and clinical therapy of Mycobacterium tuberculosis.

Description

technical field [0001] The invention belongs to the field of biomedical testing, and relates to a microbial molecular biology detection method, in particular to a method for detecting mycobacterium tuberculosis by using an aptamer technology. Background technique [0002] Tuberculosis is an infectious disease controlled by WHO, and it is still a public health problem that seriously endangers people's health. According to relevant statistics, about 1 / 3 of the world's population is infected with tuberculosis, and there are about 20 million tuberculosis patients, with 8 to 10 million new cases every year, and about 2 million people die of tuberculosis every year. The total number of deaths. The tuberculosis epidemic in my country is extremely severe. There are currently 500 million people infected with tuberculosis, accounting for 1 / 4 of the world's total. Every year, 130,000 people die from tuberculosis, which is twice the total number of deaths from other infectious and para...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04G01N33/569
Inventor 胡忠义
Owner SHANGHAI PULMONARY HOSPITAL
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