Real-time fluorescent quantitative RT-PCR kit for detection of chicken astrovirus and application thereof

Abstract

The invention discloses a real-time fluorescent quantitative RT-PCR kit for detection of chicken astrovirus and application thereof, and belongs to the field of biotechnology. The kit can achieve accurate quantitative detection of amount of mRNA expression of CAstV in a sample by reverse transcription (RT) of the mRNA sample to obtain cDNA and combination of real-time quantitative PCR detection technology. The kit can be used for the expression analysis of the CAstV in chicken embryos, and can also be used for detecting the nucleic acid level of gene of the CAstV virus in clinical cases. A rapid, accurate and reproducible detection method for detecting the level of CAstV virus infection in clinical samples is provided.

Description

technical field [0001] The invention belongs to the field of biological technology and relates to the technology of quantitative RT-PCR detection kit. Background technique [0002] Avian Astroviruses belong to the Picornaviridae family and are non-enveloped single-stranded positive-sense RNA viruses. It was first discovered in the feces of children with diarrhea in 1975. The International Committee on Taxonomy of Viruses (ICTV) classified the family Astroviridae into the genus Mamastrovirus according to the host range of virus infection. and avian Astrovirus (Avastrovirus), of which the avian Astrovirus is divided into three species, namely, avian Astrovirus GI.A type, avian Astrovirus GI.B type and avian Astrovirus GII.A type . Avian Astrovirus GI.A includes Turkey Astrovirus Type 1 (TAstV-1), and Avian Astrovirus GI.B includes Avian Nephritis Virus Type 1 (ANV-1), Avian Nephritis Virus Type 2 (ANV- 2) and Chicken Astrovirus (CAstV). The avian astrovirus GII.A type incl...

AI Technical Summary

Problems solved by technology

[0006] Before the real-time fluorescent quantitative reverse transcription polymerase chain reaction (Real-time PCR) technology appeared, whether it was direct PCR or competitive PCR, the quantification of PCR templates basically had to go through electrophoresis of PCR products, and then the electrophoresis results were analyzed by computer. Image processing, according to the brightness of the electrophoresis band to determine the amount of the final PCR product, or detect the labeled PCR product by ELISA, and then infer the amount of the starting template, but these methods are actually semi-autonomous. Quantitative level, because even if the PCR conditions are optimized, the instability of electrophoresis and subsequent steps will still affect the analysis of the results, thus affecting the purpose of quantification

[0007] At present, the research on CAstV is still in the initial stage, and various methods for detecting the expression of CAstV are still lacking.

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