Highly effective detection method for mycobacterium tuberculosis complex in clinic sample

Abstract

Sputum is a main sample for clinical test of tuberculosis, but the prior sputum processing method kills mycobacteria to different degrees during the processing process, thereby detectable rate of the tuberculosis mycobacteria is severely reduced. The invention firstly provides a method for quick and effective liquefaction of the sputum, has the characteristics of simple operational procedures and good liquefaction effect, and has the advantage that the method has no killing function on the mycobacteria in the sputum during the liquefaction process. The invention also provides a test method for tuberculosis mycobacterium complex nucleic acids with high sensitivity and strong specificity, wherein, specific amplified products of Ag85B gene are tested through fluorescent PCR amplification method and specific primers and a Taqman fluorescent probe are designed according to a tuberculosis mycobacterium complex Ag85B gene order, thereby the tuberculosis mycobacterium complex is tested. The invention is implemented by the two mutually independent and mutually associated inventions, and provides a method for testing the tuberculosis mycobacterium complex in clinical samples, which has the advantages of high efficiency, quickness, high sensitivity and strong specificity.

Description

technical field [0001] The invention relates to the technical field of clinical microbial nucleic acid diagnosis, in particular to a technology for detecting Mycobacterium tuberculosis nucleic acid in clinical samples with high sensitivity, strong specificity and high efficiency. No killing effect on mycobacteria in sputum. Background technique [0002] Mycobacteriaceae includes only one genus of Mycobacterium, which is divided into Mycobacterium tuberculosis complex (MTBC) and nontuberculosis mycobacterium (nontuberculosis mycobacterium, NTM); Mycobacterium tuberculosis complex includes Mycobacterium tuberculosis (M.tuberculosis, MTB), Mycobacterium bovis (M.bovis), Mycobacterium Africanum (M.africanum) and Mycobacterium microti (M.micoti), Mycobacterium tuberculosis Mycobacterium Wagyu is the pathogen that causes tuberculosis in humans, and Mycobacterium africanum is the pathogen that causes tuberculosis in tropical countries in Africa. Mycobacterium tuberculosis can exi...

AI Technical Summary

Problems solved by technology

[0008] Second, sputum smears, radiographs, and culture tests may not accurately identify active TB disease, especially in HIV-positive patients

[0009] Third, sputum smear examination, X-ray imaging, and culture tests may not be able to strictly distinguish latent and active tuberculosis, as well as the sensitivity and resistance of the disease to drugs

The copy number of 16srRNA in the cell is about 1000-10000copies / cell, so AMTD2 detection sensitivity is extremely high, and it is certified by FDA to detect smear-positive and smear-negative specimens; its main disadvantage is that there is no internal standard, and it cannot be amplified Detection of Response Inhibition

At the same time, the above two products are expensive, and the liquefaction method used is the NaOH-NALC method, which can kill Mycobacterium tuberculosis in sputum samples to a certain extent, so it is difficult to detect Mycobacterium tuberculosis in sputum of patients with latent infection

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