CSFV (classical swine fever virus), PCV2 (porcine circovirus type 2) and PRV (pseudorabies virus) multiple SYBR Green I real-time fluorescence PCR (polymerase chain reaction) primers and detection method thereof

Abstract

The invention discloses CSFV (classical swine fever virus), PCV2 (porcine circovirus type 2) and PRV (pseudorabies virus) multiple SYBR Green I real-time fluorescence PCR (polymerase chain reaction) primers and a detection method thereof. The CSFV primers have the following sequences: the upstream primer P1: 5'-AAACGGAGGGACTAGCCGT-3', and the downstream primer P2: 5'-TGCCATGTACAGCAGAGA-3'; the PRV primers have the following sequences: the upstream primer P3: 5'-CTCGCCATCGTCAGCAA-3', and the downstream primer P4: 5'-GCTGCTCCTCCATGTCCTT-3'; and the PCV2 primers have the following sequences: theupstream primer P5: 5'-GGGCCAGAATTCAACCTTACCC-3' and the downstream primer P6: 5'-CGCACCTTCGGATATACTGTCA-3'. Through a CSFV, PCV2 and PRV triple SYBR Green I quantitative PCR detection method, CSFV, PRV and PCV2 three types of DNA viruses can be simultaneously detected, and at least 183 copies of CSFV, 231 copies of PRV and 203 copies of PCV2 can be detected. In the invention, a specificity test result is good, only three specificity peak values appear, repeated tests have good stability, and a novel quick, sensitive, specific and accurate method with low cost is provided for effectively detecting and identifying mixed infection, and is conductive to identifying piggysow reproductive obstacle viruses.

Description

technical field [0001] The invention relates to a detection method of porcine virus, in particular to CSFV, PCV2 and PRV multiple SYBR Green I real-time fluorescent PCR primers and detection method. Background technique [0002] Swine fever virus (CSFV), porcine pseudorabies virus (PRV) and porcine circovirus type 2 (PCV2) can all cause pregnancy to varying degrees and cause infertility in breeding pigs, that is, boar testicle swelling, semen quality decline, and loss of breeding ability ; Sows show anoestrus, return to estrus, infertility; pregnant sows have miscarriage, premature birth, stillbirth, mummies, weak piglets and newborn piglets are sick and cause a large number of deaths, etc., causing huge economic losses to the pig industry. Effective control of such infectious diseases is one of the key factors to ensure the healthy development of the pig industry. [0003] At present, there have been a lot of related research on the above three viruses at home and abroad, ...

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is that the PCR detection method of CSFV, PCV2 and PRV is prone to false negative results, and cannot be quantified. A kind of CSFV, PCV2 and PRV multiple SYBR Green I real-time fluorescent PCR primers and detection method are provided

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