Fluorescent PCR reaction liquid and kit for mycobacterium parting identification

Abstract

The invention discloses a fluorescent PCR reaction liquid for mycobacterium parting identification. The fluorescent PCR reaction liquid contains amplification primers and a molecular beacon probe. Specifically, the molecular beacon probe includes at least one of 12 mycobacterial molecular beacon probes with both ends provided with fluorescent groups and quenching groups respectively, and the sequences are shown as SEQ ID NO.1-SEQ ID NO.12. The invention also discloses a kit having the PCR reaction liquid. With the fluorescent PCR reaction liquid and kit provided by the invention, 12 mycobacteria including mycobacterium tuberculosis and nontuberculosis mycobacteria can be quickly and accurately detected and identified.

Description

technical field [0001] The invention relates to a fluorescent PCR reaction solution and a kit for mycobacteria typing and identification. Background technique [0002] Mycobacteria include Mycobacterium tuberculosis (M. tuberculosis, MTB), nontuberculous mycobacteria (Nontuberculous Mycobacteria, NTM) and Mycobacterium leprae. NTM often exists in the natural environment, and more than 150 species have been discovered so far, about 1 / 3 of which are related to human diseases. [0003] Previous tuberculosis epidemiological data in my country show that the NTM isolation rate rose from 4.9% in 1990 to 11.1% in 2000 and 22.9% in 2010, which basically reflects the obvious upward trend of NTM disease in my country. [0004] Clinical studies have found that NTM is resistant to most anti-mycobacterial drugs. Therefore, the principle of drug use for NTM disease is significantly different from that of tuberculosis, and the types and course of drug use for different types of NTM disease...

AI Technical Summary

Problems solved by technology

[0005] However, the pathological findings of NTM disease are difficult to distinguish from tuberculosis. Traditional clinics can only rely on the growth characteristics of mycobacteria, pigments, growth speed, colony morphology and parameters such as biochemical tests to make identification. This method is time-consuming, laborious, and There is no standardized reagent, lack of sensitivity and specificity, making many results difficult to judge

The determination of mycolic acids by high performance liquid chromatography and the determination of fatty acids by gas chromatography have limitations in the identification of strains, which are affected by the growth environment and nutritional status of the strains, and require special instruments

[0006] Genetic detection methods can detect different types of NTM, but the direct sequence determination of PCR products and restriction endonuclease mapping are complex operations and are not suitable for detection of mixed infections

Ordinary fluorescent quantitative PCR can only distinguish NTM and MTC (Mycobacterium tuberculosis), but cannot type NTM

Gene chips and reverse dot hybridization can be used to type NTM, but both methods are complicated to operate and are not suitable for large-scale clinical use

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