Fluorescent quantitative RT-PCR detection method of M.tuberculosis-complex

A technique for Mycobacterium tuberculosis and fluorescence quantification, which is applied in the determination/inspection of microorganisms, fluorescence/phosphorescence, biochemical equipment and methods, etc. Reliable source, avoid use, avoid contamination effect
CN101974630AInactive Publication Date: 2011-02-16FUDAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
FUDAN UNIV
Publication Date
2011-02-16
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the technical field of biological engineering detection, in particular relates to a fluorescent quantitative RT-PCR detection method of M.tuberculosis-complex. The invention includes the following steps: M.tuberculosis-complex specificity quantitative RT-PCR primer and probe are designed; standard molecule is constructed; coextraction of bacterium samples RNA and DNA is carried out; RT-PCR and PCR are respectively carried out on the same sample; and a fluorescent quantitative PCR detection method is created and optimized. By applying the method of the invention to detect M.tuberculosis-complex, the advantages of fast speed, high sensitivity and strong specificity can be achieved, dead bacterium and living bacterium can be distinguished, and the defects that detection period is long, positive rate is low and distinguishing of dead bacterium and living bacterium can not be realized in the existing detection method can be overcome, thus having important practical application value.
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Description

technical field

[0001] The invention belongs to the technical field of bioengineering detection, in particular to a fluorescence quantitative detection method for mycobacterium tuberculosis complex in clinical samples. Background technique

[0002] The Mycobacterium tuberculosis complex includes Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum, and Mycobacterium microti. Mycobacterium tuberculosis is the main pathogen that causes human tuberculosis. In addition, Mycobacterium bovis not only causes bovine tuberculosis, but also a small number of human tuberculosis. Mycobacterium africanum is weakly pathogenic and is the causative agent of tuberculosis in tropical Africans. Mycobacterium microti is not pathogenic to humans.

[0003] Tuberculosis is one of the public health problems that seriously endanger the world. Since the 1990s, tuberculosis has "resurrected" around the world. Many countries, including countries with better tuberculosis epidemic c...

Claims

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