Tubercle bacillus drug tolerance detection reagent kit and tubercle bacillus drug tolerance detection method

Abstract

The invention provides a tubercle bacillus drug tolerance detection reagent kit and method. The tubercle bacillus drug tolerance detection reagent kit comprises a tubercle bacillus drug tolerance detection reagent, wherein the tubercle bacillus drug tolerance detection reagent comprises a sequencing primer in accordance with a tubercle bacillus drug tolerance gene; the tubercle bacillus drug tolerance gene comprises one or more genes of rpoB, katG, inhA-promoter, inhA-structural, furA, embB, ubiA, pncA, rpsA, gyrA, gyrB, eis, rpsL, rrs, tlyA, rplC and rrl; and further, the reagent kit also contains a tubercle bacillus nucleic acid detection reagent, and the tubercle bacillus nucleic acid detection reagent comprises a primer pair 1 in accordance with IS6110, a primer pair 2 in accordance with the IS6110 and a probe primer in accordance with the IS6110. Through the adoption of the tubercle bacillus drug tolerance detection reagent kit disclosed by the invention, tubercle bacillus nucleicacid in samples can be quickly detected, and positive samples can be further subjected to drug tolerance detection; and the tubercle bacillus drug tolerance detection reagent kit has good sensitivity, good specificity and good accuracy, and can perform mutation detection on 48 sites of 17 drug tolerance genes of common antituberculosis drugs and fragment deficiency detection of an intergenic region, so that the tuberculosis medication can be more accurately and comprehensively guided.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, and in particular relates to a drug resistance detection kit of Mycobacterium tuberculosis and a method for detecting drug resistance of Mycobacterium tuberculosis. Background technique [0002] Mycobacterium tuberculosis, commonly known as Mycobacterium tuberculosis, is the pathogenic bacteria that causes tuberculosis. Mycobacterium tuberculosis is a slender and slightly curved bacillus that can invade susceptible organisms through respiratory tract, digestive tract or skin damage, causing tuberculosis in various tissues and organs, among which pulmonary tuberculosis is the most common through the respiratory tract. Drug-resistant TB occurs when the Mycobacterium tuberculosis infected with the patient becomes resistant to one or more anti-TB drugs. According to the report of WHO in 2008, the total drug resistance rate of tuberculosis in the world is 20.0%, and the rate of multidr...

AI Technical Summary

Problems solved by technology

First, AFB smears have low sensitivity (30-40%) and their accuracy is often limited

Second, while culture-based methods have high accuracy and have long been considered the gold standard for laboratory TB diagnosis, the long turnaround time (3-8 weeks) for mycobacterial cultures often leads to delays in diagnosis

The reverse dot hybridization method for molecular detection has low sensitivity, complex operation and unreliable results

Commercially available real-time polymerase chain reaction (PCR) assays such as Abbott Real-Time MTB Assay and GeneXpert MTB / RIF Ultra provide timely and accurate TB diagnosis but are expensive

However, most of these methods are used to detect Mycobacterium checkeriensis. GeneXpert MTB / RIF can detect drug-resistant tuberculosis, but it can only detect a drug-resistant gene of rifampicin, which leads to patients who are treated with other drugs. Drug resistance or other resistance genes to rifampicin drug resistance cannot be detected

Molecular reverse dot hybridization can also be detected, but currently only ten sites of five drug-resistant genes of four drugs (isoniazid, rifampicin, ethambutol, streptomycin) can be detected. Resistance genes beyond four drugs remain undetectable

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