TaqMan probe-based Senecavirus A fluorescent quantitative RT-PCR (reverse transcription-polymerase chain reaction) detecting method and kit

Abstract

The invention discloses a TaqMan probe-based Senecavirus A fluorescent quantitative RT-PCR detecting method and kit. The sequences of primers applied in the TaqMan probe-based Senecavirus A fluorescent quantitative RT-PCR detecting method are shown as SEQ ID NO.3 and SEQ ID NO.4, and the sequence of a probe applied in the TaqMan probe-based Senecavirus A fluorescent quantitative RT-PCR detecting method is shown as SEQ ID NO.5. The primers and the probe are obtained by performing sequence comparison and analysis according to the VP1 gene sequences of 41 SVA (Senecavirus A) strains epidemic in China since 2015 which are published in NCBI (National Center for Biotechnology Information) GenBank databases, and performing design within the conserved regions of the sequences; the sequences of theprimers and the probe are high in conservatism among analyzed virus strains. The TaqMan probe-based Senecavirus A fluorescent quantitative RT-PCR detecting method is high in specificity, sensitivityand stability, and can be successfully applied to clinical SVA inflection identification and diagnosis and provide good technical support for rapid swine vesicular disease identification and diagnosisin China.

Description

technical field [0001] The invention relates to a type A Seneca virus fluorescence quantitative PCR detection primer, a probe, a detection method and a kit based on a TaqMan probe method, and belongs to the technical field of bioengineering. Background technique [0002] Seneca Valley virus (SVV), also known as type A Seneca virus (SenecavirusA, SVA), was initially used as a contaminant in the culture of human embryonic kidney cells (PER. Identification for the first time. Existing studies have shown that SVA is also one of the important pathogens causing porcine idiopathic vesicular disease (PIVD), and can lead to high mortality in piglets. SVA belongs to Picornaviridae and Senecavirus. Virus particles have no envelope structure and are single-stranded positive-strand RNA viruses. The genome size is about 7.3kb, including the untranslated regions (Untranslated regions) at both ends. , UTRs) and a large open reading frame (Open reading frame, ORF), encoding four structural...

AI Technical Summary

Problems solved by technology

Most of the existing qRT-PCR detection methods in my country are established based on the genome sequence of the earliest isolated strain SVV-001 in the United States in 2002, and there is a lack of data support for clinical sample detection, and the detection effect on the current new epidemic strains is uncertain

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