Haemophilus paragallinarum fluorogenic quantitative PCR detection method

Abstract

The invention discloses a haemophilus paragallinarum fluorogenic quantitative PCR detection method. According to the haemophilus paragallinarum fluorogenic quantitative PCR detection method, a hagA gene with the high degree of specificity and conservative property is selected as an amplification target gene, SYBR Green I fluorochrome is taken as a detection target, through the continuous optimization of fluorogenic quantitative PCR reaction conditions, the preparation of a standard substance and the establishment of a standard curve, the haemophilus paragallinarum fluorogenic quantitative PCR detection method of the SYBR Green I fluorochrome jogged with DNA, and the DNA of a haemophilus paragallinarum hagA genetic template can be quantified accurately. The haemophilus paragallinarum fluorogenic quantitative PCR detection method has the advantages of being strong in specificity, high in sensibility, simple in operation, rapid in detection, accurate in quantification and the like, the haemophilus paragallinarum fluorogenic quantitative PCR detection method can be used for clinic rapid diagnosis of haemophilus paragallinarum and the quantitative analysis of the degree of infection by the haemophilus paragallinarum, and the haemophilus paragallinarum fluorogenic quantitative PCR detection method is suitable for popularization and application.

Description

technical field [0001] The invention relates to a method for detecting Haemophilus paragallinarum. Specifically, the present invention relates to a fluorescent quantitative PCR detection method for detecting Haemophilus paragallinarum. Background technique [0002] Haemophilus paragallinarum (H.gallinarum), also known as Avibacterium paragallinarum Apg, is the pathogen that causes infectious coryza IC in chickens. It mainly causes acute upper respiratory tract symptoms in chickens, mainly manifested as nasal passages. 1. A large amount of serous or mucous secretions appear in the sinuses, facial edema and conjunctivitis, which can lead to a decrease in the laying rate of laying hens (10%-40%), poor growth of broilers, and increased elimination rate. caused huge economic losses. [0003] At present, the laboratory diagnosis methods of Haemophilus paragallinarum mainly include the isolation and identification of bacteria, animal regression test and conventional PCR method. ...

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Problems solved by technology

[0002] Haemophilus paragallinarum (H.gallinarum), also known as Avibacterium paragallinarum Apg, is the pathogen that causes infectious coryza IC in chickens. It mainly causes acute upper respiratory tract symptoms in chickens, mainly manifested as nasal passages. 1. A large amount of serous or mucous secretions appear in the sinuses, facial edema and conjunctivitis, which can lead to a decrease in the laying rate of laying hens (10%-40%), poor growth of broilers, and increased elimination rate. caused huge economic losses

[0003] At present, the laboratory diagnosis methods of Haemophilus paragallinarum mainly include the isolation and identification of bacteria, animal regression test and conventional PCR method. Bacillus is a delicate bacterium, if the storage of diseased materials, poor transportation conditions or poor quality of medium, will lead to the failure of the test

Although the conventional PCR diagnostic method has improved the sensitivity of detecting Avibacterium paragallinarum from diseased materials and isolated suspected cultures, there are problems such as false positives and inaccurate quantification, and the nucleic acid dye ethidium bromide used in the electrophoresis process, Harmful to the experimental environment and experimenters

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