Fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) kit for detecting influenza A virus subtype H7N9

Abstract

The invention provides a fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR) kit for detecting an influenza A virus subtype H7N9. The fluorescent quantitative RT-PCR kit can be used for detection of influenza A viruses and the influenza A virus subtype H7N9. The fluorescent quantitative RT-PCR kit comprises a quantitative RT-PCR reaction solution, an enzyme mixed liquor, a primer and probe mixed liquor, standard substances of influenza A viruses, H7, N9 and RNaseP, positive reference substances of influenza A viruses, H7, N9 and RNaseP), and negative reference substances. Specific primers and probes are designed according to conserved sequences of influenza A viruses, H7 and N9. The RNaseP primers and probes are used as internal references. Through the one-step quadruple real-time fluorescent RT-PCR technology, the influenza A virus and the influenza A virus subtype H7N9 in the sample can be fast and accurately detected. The fluorescent quantitative RT-PCR kit has a reasonable design, very high singularity, sensitivity and repeatability, can be used for laboratory emergency diagnosis and fast screening of an epidemic disease caused by the influenza A virus subtype H7N9, and for an epidemiology study on the influenza A virus and the influenza A virus subtype H7N9 causing fever and respiratory tract syndrome.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a fluorescent quantitative RT-PCR detection kit, in particular to a one-step quadruple real-time fluorescent quantitative RT-PCR for detecting nasopharyngeal swabs, sputum, and nasopharyngeal aspiration of patients in the same reaction tube. The detection method of influenza A virus and influenza A virus H7N9 subtype nucleic acid in specimens such as animals can be applied to laboratory emergency diagnosis, rapid screening, clinical diagnosis and influenza A in patients with febrile respiratory syndrome caused by outbreaks of H7N9 avian influenza. Epidemiological study of influenza virus and influenza A virus H7N9 subtype. Background technique [0002] Influenza A virus H7N9 subtype is the first new subtype influenza virus discovered in the world. It was first discovered in Shanghai and Anhui in March 2013. The H7N9 subtype is a type of influenza A, which was only found in pou...

AI Technical Summary

Problems solved by technology

Due to the high conditions and technical requirements for pathogen isolation and culture, and the long period; and the H7N9 subtype of influenza A virus, as a new subtype of influenza virus discovered, lacks antigens and antibodies, and there is no mature rapid diagnostic reagent on the market; real-time fluorescent quantitative PCR technology and Its fully enclosed single-tube amplification, simple and quick, good repeatability, real-time quantification, less pollution and other advantages overcomes the shortcomings of previous clinical diagnosis, has high specificity and sensitivity, and provides an accurate and reliable laboratory for clinical diagnosis. It can be used as an effective and rapid detection method for clinical pathogen diagnosis

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