Method for detecting drug resistance of antituberculosis drugs based on overlap-extension PCR

Abstract

The invention relates to a method for detecting the drug resistance of antituberculosis drugs based on an overlap-extension PCR. By adopting an overlap-extension PCR method, four drug-resistance related segments of rpoB, embB and katG genes and an inhA promoter are connected and amplified into a fusion segment with the length of about 700 bp in a single tube, drug-resistance mutation information of the four segments is obtained just through a one-time sequencing reaction, and accordingly the condition of drug resistance of some kind of mycobacterium tuberculosis to three kinds of first-line antituberculosis drugs is known. If the method is applied and popularized clinically, the detection cost is greatly reduced, the economic burden of a patient is reduced, and the good social benefit is achieved. In short, by means of the method that the single-tube PCR amplification rpoB-embB-katG-inhA fusion segment is used for sequencing, so that the drug resistance condition of three kinds of first-line antituberculosis drugs is detected, the economic burden is reduced for detection of clinical drug resistance tuberculosis, the detection accuracy is improved, and the method is easy and convenient to implement, accurate and low in cost.

Description

Technical field [0001] The present invention belongs to the field of gene technology, and relates to a method for detecting the resistance of three anti-tuberculosis drugs at a time based on overlap extension PCR. Background technique [0002] Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (Mtb). It is a major public health problem that seriously endangers human health. In recent years, the drug resistance of Mycobacterium tuberculosis has gradually deteriorated, and the epidemic situation of drug-resistant tuberculosis is very serious, so the importance of diagnosis of drug-resistant tuberculosis has become increasingly prominent. Although the culture-based method is still the gold standard for the diagnosis of drug-resistant tuberculosis, the traditional drug susceptibility test to detect Mtb drug resistance takes up to several weeks, which delays the treatment of patients and may lead to the emergence of new drug resistance, which is lab...

AI Technical Summary

Problems solved by technology

[0005] In summary, these genotypic drug resistance detection methods have the advantages of high efficiency and rapidity, and can quickly diagnose tuberculosis and some drug resistance, but because of the relatively high cost of detection

Expensive prices increase the financial burden on patients in poor areas

There are still a small number of cases where the test results are not consistent with the sequencing results. In the case of synonymous mutations in the probe binding sites in the coding regions of drug resistance-related genes, false false drug resistance results appear.

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