Method and kit for detecting drug resistance of mycobacterium tuberculosis

Abstract

The invention discloses a method and a kit for detecting drug resistance of mycobacterium tuberculosis. The invention provides special-purpose primers and special-purpose probes for identification of drug resistance of mycobacterium tuberculosis in a sample needing to be detected, wherein the special-purpose primers comprise multiple primer groups; each one of the primer groups comprises a common primer and multiple specific primers for amplification of alleles corresponding to all mutational sites of same-purpose genes; the special-purpose probes comprise multiple types of probes; and each one of the probes specifically binds with target sequences corresponding to the common primer and the specific primers of the primer group. Experiments of the invention prove that the special-purpose primers and the special-purpose probes can be combined by an efficient and sensitive duplex Taqman real-time fluorescent polymerase chain reaction (PCR) technology and a sequence specific primer (SSP) amplification technology so that drug resistance of mycobacterium tuberculosis in a detected sample can be detected in a short time.

Description

technical field [0001] The invention relates to a method and a kit for detecting drug resistance of mycobacterium tuberculosis. Background technique [0002] Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (MTB). According to the data provided by the World Health Organization (WHO), the situation of tuberculosis in the world in 2002 was: about 3 billion infected people and 8.8 million new tuberculosis cases. Among them, there were 3.9 million new smear-positive tuberculosis cases, an increase of 1.1% compared with 2001, and 3.2% of new cases were multi-drug resistant, and the death toll of tuberculosis reached 1.8 million. Retrospective survey results show that the total drug resistance rate of MTB in China is 27.8%, the initial drug resistance rate and acquired drug resistance rate are 18.6% and 46.5%, respectively, and the multidrug resistance rate is 10.7%, of which initial and acquired multidrug resistance The rates were 7.6% and 17.1%, respe...

AI Technical Summary

Problems solved by technology

[0009] (1) DNA sequencing method (DNA sequencing) directly sequences the PCR product of the drug-resistant gene fragment, and then compares it with the same DNA fragment of the standard sensitive strain to analyze the position and distribution of the base mutation, but this method is cumbersome, and the Most of the equipment that needs to be concentrated in some large sequencing companies is difficult to popularize

[0010] (2) PCR-SSCP method PCR-SSCP, that is, polymerase chain reaction-single-strand conformational polymorphism (polymerasechain reaction-singlestranded conforma-tional polymorphism, PCR-SSCP), but this method detects DNA fragments with single base changes The length should be less than 200bp, and relatively complete drug resistance information cannot be obtained from small fragments, and the sensitivity decreases with the increase of DNA length

This method has high specificity, but can only analyze gene mutations at specific sites in known sequences

[0012] (4) Line probe assay (LiPA) LiPA is expensive and difficult to popularize

[0013] (5) Although gene chip technology has made great progress, there are still defects, such as the possibility of wrong nucleotides being mixed in when synthesizing probes

In addition, its operation process is complicated and the required materials are expensive, which hinders the promotion of this technology.

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