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Method for extracting live bacteria RNA in Mycobacterium tuberculosis and detection kit thereof

A technology of Mycobacterium tuberculosis and a detection kit, which is applied in the biological field, can solve the problems of complex production and limited wide application, and achieve the effects of improving the detection sensitivity and specificity and reducing the false positive rate.

Inactive Publication Date: 2010-06-30
SHANGHAI FOSUN PHARMA (GROUP) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this technology requires specific antibodies, and the production is relatively complicated
At the same time, false positives limit the widespread application of this technique

Method used

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  • Method for extracting live bacteria RNA in Mycobacterium tuberculosis and detection kit thereof
  • Method for extracting live bacteria RNA in Mycobacterium tuberculosis and detection kit thereof
  • Method for extracting live bacteria RNA in Mycobacterium tuberculosis and detection kit thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0076] Detection of Mycobacterium tuberculosis Viability by Real-time Fluorescent Quantitative RT-PCR and Its Application

[0077] 1. Primer and probe design and synthesis:

[0078] Using the full-length cDNA sequence of Mycobacterium tuberculosis (GenBank accession number NC_000962) as a template, Oligo software was used to analyze the positions of TaqMan primers and probes, and the best combination was selected based on the consideration of the genomic DNA sequence of Mycobacterium tuberculosis.

[0079] The upstream primer sequence of reference product PCR is: 5'-CTG CGG TTT ATC TGC TCG AC-3'; the downstream primer sequence is: 5'-GAC AGC GAG CCG GCG TAG A-3';

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Abstract

The invention relates to a method for extracting live bacteria RNA in Mycobacterium tuberculosis and a detection kit thereof, and particularly provides a method for extracting live bacteria RNA in Mycobacterium tuberculosis and a detection kit which is used for the live bacteria RNA in the Mycobacterium tuberculosis and is obtained by applying the method and combining fluorescent quantitative PCR technology. The kit can accurately, sensitively and quickly identify dead bacteria and the live bacteria of the Mycobacterium tuberculosis, reduce the cost and shorten the detection time, and more importantly, the kit is the basis of studies such as the diagnosis of tuberculosis, medicinal susceptibility experiments, the monitoring of chemotherapy response, the screening of new antitubercular medicaments, the prevention of the tuberculosis and the like.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a fluorescent quantitative RT-PCR kit for quantitatively detecting live bacteria of Mycobacterium tuberculosis. Combined with the real-time fluorescent quantitative PCR detection technology, the viable bacteria of Mycobacterium tuberculosis in the specimen to be tested can be accurately quantified. Background technique [0002] Since Koch discovered that Mycobacterium tuberculosis is the pathogenic bacterium of tuberculosis in 1882, for more than 100 years, human beings have made great progress in the biological characteristics, pathogenic mechanism, drug resistance mechanism, diagnostic methods, and development of anti-tuberculosis drugs of Mycobacterium tuberculosis. Huge achievement. Today, tuberculosis is still one of the most widespread infectious diseases affecting human health and the highest fatality rate. Seeking rapid and accurate diagnostic methods ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/10
Inventor 沈维祥吴大治夏懿
Owner SHANGHAI FOSUN PHARMA (GROUP) CO LTD
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