Development and verification of a duplex real-time PCR for detection of bovine viral diarrhea virus and African swine fever virus in pig vaccines
10.13200/j.cnki.cjb.003911
- VernacularTitle:猪用疫苗中牛病毒性腹泻病毒和非洲猪瘟病毒双重荧光PCR检测方法的建立及验证
- Author:
YU Xinyou
- Publication Type:Journal Article
- Keywords:
Bovine viral diarrhea virus(BVDV);
African swine fever virus(ASFV);
Real-time PCR;
TaqMan probe
- From:
Chinese Journal of Biologicals
2024;37(9):1122-1126+1132
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop and verify a duplex real-time PCR method for simultaneous detection of bovine viral diarrhea virus(BVDV) and African swine fever virus(ASFV) in pig vaccines and apply it preliminarily.Methods According to the sequence of BVDV 5'UTR and ASFV VP72 included in NCBI,a pair of specific primers and a TaqMan probe were designed respectively,and a duplex real-time PCR method for simultaneous detection of BVDV and ASFV was established. The developed method were verified for the sensitivity,specificity and precision,and used to detect BVDV and ASFV of a total of 76 batches of live vaccines against porcine pseudorabies virus(PRV)and classical swine fever virus(CSFV),as well as live vaccines against porcine PRV added with BVDV and recombinant plasmid pMD-VP72,5 bottles for each,respectively. The BVDV and ASFV were rechecked and detected using the methods stipulated in the third part of Veterinary Pharmacopoeia of the People's Republic of China(2020 edition)and Announcement No. 172 of the Ministry of Agriculture and Rural Affairs of the People's Republic of China.Results 4. 2 copies/μL of BVDV and 8. 7 copies/μL of ASFV recombinant plasmids were detected by the developed duplex real-time PCR;No cross reaction with other common pathogens was observed and the coefficient of variation(CV)of repeatability and precision was not more than 2%.Conclusion The duplex real-time PCR method developed in this experiment showed good sensitivity,specificity and precision,which might be used for the rapid detection of exogenous viruses in pig vaccines and the quality control of pig vaccines and other biological products.
