Reagent kit and method for detecting M.tuberculosis isoniazide drug tolerance mutation gene

Abstract

The invention provides a reagent kit and method for detecting an M.tuberculosis isoniazide drug tolerance mutation gene, particularly a multiplex PCR amplification system of three target genes katG, inhA and aphC with M.tuberculosis isoniazide drug tolerance mutation is established, and through a high-resolution melting curve method, wild types and different mutation types can be distinguished. Inaddition, through binding a molecular beacon probe, a multiple asymmetric PCR reaction can be realized, so that the mutation site of a tuberculosis resisting bacterial strain can be detected quicklyand accurately, and multiple sites can also be detected in a disposable manner.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a kit and a method for detecting the isoniazid resistance mutation gene of mycobacterium tuberculosis. Background technique [0002] Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis, which is characterized by a long incubation period and a high fatality rate. So far, it is still one of the three major infectious diseases that seriously endanger human health worldwide, along with AIDS and malaria. Mycobacterium tuberculosis, which belongs to a class of pathogenic bacteria in the genus Mycobacterium, is the pathogen of most tuberculosis. At present, about 30% of people in the world have been infected with tuberculosis, and the annual death toll is nearly 3 million. Therefore, tuberculosis has always been It is a global health problem that threatens human health. Since the 1940s and 1950s, Mycobacterium tuberculosis has also been effecti...

AI Technical Summary

Problems solved by technology

[0005] Among the phenotypic detection methods, the traditional Roche medium drug susceptibility detection method and the BACTEC MGIT 960 system detection method are cumbersome to operate and the cycle is long; the DNA sequencing method in the molecular biology detection method is expensive and difficult to popularize, and it is difficult to implement the linear hybridization method. Meet the requirements of clinical rapid detection, and although the gene chip method has made some progress, there may be wrong nucleotides mixed in when synthesizing probes

And the operation process is complicated, the material is expensive, the cost is high, and it is also difficult to popularize

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