CSFV (Classical Swine Fever Virus), PRRSV (Porcine Reproductive and Respiratory Syndrome Virus), PCV2 (Porcine Circovirus 2) and SIV (Swine Influenza Virus) H9 subtype multiplex SYBR Green I real-time fluorescence PCR (Polymerase Chain Reaction) primer and detection method thereof

Abstract

The invention discloses a CSFV (Classical Swine Fever Virus), PRRSV (Porcine Reproductive and Respiratory Syndrome Virus), PCV2 (Porcine Circovirus 2) and SIV (Swine Influenza Virus) H9 subtype multiplex SYBR Green I real-time fluorescence PCR (Polymerase Chain Reaction) primer and a detection method thereof. The sequences of CSFV primers are as follows: an upstream primer P1: 5'-AAACGGAGGGACTAGCCGT-3', and a downstream primer P2: 5'-TGCCATGTACAGCAGAGA-3'; the sequences of PRRSV primers are as follows: an upstream primer P3: 5'-AAACCAGTCCAGAGGCAAG-3', and a downstream primer P4: 5'-CCACAGTGTAACTTATCCTC-3'; the sequences of PCV2 primers are as follows: an upstream primer P5: 5'-GGGCCAGAATTCAACCTTACCC-3', and a downstream primer P6: 5'-CGCACCTTCGGATATACTGTCA-3'; and the sequences of SIV H9 subtype primers are as follows: an upstream primer P7: 5'-GGAACAACGCTTACCCTG-3', and a downstream primer P8: 5'-GAGACCATTGACAAGAGGC-3'. In the invention, specific primers for amplifying CSFV, PRRSV, PCV2 and SIV H9 subtype are designed respectively, and the CSFV, PRRSV, PCV2 and SIV H9 subtype can be detected simultaneously with a quadruple SYBR Green I real-time quantitative PCR detection method. The method has high specificity, repeatability and sensitivity, a novel method for detecting the CSFV, PRRSV, PCV2 and SIV H9 subtype is provided and can be taken as a purified detection method for the CSFV, PRRSV, PCV2 and SIVH9 subtype in a large scale pig farm, and a swine herd without CSFV, PRRSV, PCV2 or SIV H9 subtype is established.

Description

technical field [0001] The invention relates to a virus detection method, in particular to CSFV, PRRSV, PCV2 and SIV H9 subtype multiple SYBR Green I real-time fluorescent PCR primers and a detection method thereof. Background technique [0002] Classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) can cause pregnancy to varying degrees and cause infertility in pigs, that is, testicular swelling in boars, decreased semen quality, loss of breeding ability; sows show no estrus, return to estrus, and infertility; pregnant sows have abortions, premature births, stillbirths, mummies, weak piglets, and newborn piglets are sick and lead to a large number of deaths, which has caused huge economic losses to the pig industry. loss. SIV can cause the occurrence and spread of zoonotic diseases, threatening human safety. Effective control of such infectious diseases is one of the key factors to ensure human ...

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the present invention is that these viruses of CSFV, PRRSV, PCV2 and SIV H9 subtypes often appear in the form of mixed infection, and it is easy to cause false negatives to diagnose by separating virus and antibody detection, and the operation is cumbersome. Multiple SYBR Green Ⅰ real-time fluorescent PCR primers and detection methods for CSFV, PRRSV, PCV2 and SIV H9 subtypes

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