Establishment method for Taqman MGB probe real-time fluorescence quantitative PCR detection system taking outer membrane protein as target gene

Abstract

The invention relates to a method for rapid detection and diagnosis of Chlamydophila psittaci by using real-time PCR which is one of emerging molecular biotechnology. Based on the conserved domain of the variable domain III (VD3) of the main outer membrane protein (MOMP) gene of the Chlamydophila psittaci, the detection system of the invention designs a pair of primers and a relevant Taqman MGB probe. The system can detect Chlamydophila psittaci genes existing in bird dung and the sensitivity of the system is obviously higher than the sensitivity of common PCR. Since the Real-time PCR requires no post-PCR processing, the cross contamination possibly caused by agarose gel electrophoresis is avoided and thereby the accuracy and the credibility of the detection are improved.

Description

Technical field: The invention belongs to the field of molecular biology, and in particular relates to a method for establishing a Taqman MGB probe real-time fluorescent quantitative PCR detection system using the outer membrane protein of chlamydia psittaci as a target gene. Background technique: Chlamydophila psittaci (Cps) is a species of the genus Chlamydia in the family Chlamydiaceae. It is a microorganism between bacteria and viruses that parasitizes in eukaryotic cells. It can cause infectious atypical pneumonia in animals and humans, so the psittacosis caused by it is listed as a second-class infectious disease by the International Office of Epizootics (OIE), and thus has attracted more and more attention from researchers around the world ; Simultaneously because this pathogen can also be used as a bacterial weapon, it is therefore also included in the list of pathogens that are prohibited from being transferred and exported without permission in the United States. ...

AI Technical Summary

Problems solved by technology

At present, although the full-length sequences of 16S and 23S rRNA genes have been successfully used as target gene sequences at home and abroad, the detection of chlamydia has been realized by using techniques such as PCR. The homology of its nucleotide sequence is very high, so it is impossible to implement rapid and accurate detection and distinguish the species, resulting in false negative or false positive, which causes considerable difficulty in the diagnosis of the pathogen

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