猪繁殖与呼吸综合征活疫苗中病毒含量荧光定量PCR测定方法的建立及初步应用

Abstract

Abstract: To establish a rapid, sensitive and specific method for evaluation of porcine reproductive and respiratory syndrome (PRRS) attenuated vaccine, two pairs of primers were designed and synthesized according to porcine reproductive and respiratory syndrome virus (PRRSV) genes in GenBank. PRRSV genes were obtained by RT-PCR and constructed recombinant plasmids pMD19-T-PRRSV. SYBR GreenⅠ method was adopted to improve the Real-time PCR and the method was tested for specificity, repeatability and stability. The results shwed that there was a good linear correlation coefficient (R²=0.9989) as Ct values ranged from 7.43×10⁰ to 7.43×10⁸ copies/μL. Using this method to test PRRS vaccines from different companies, we found that there were significant differences (47.9 times) in the levels of virus among six companies’ vaccines. The Real-time PCR for detecting PRRS vaccine virus can be used for evaluation of vaccine during production process and animals immunization.

Key words:

porcine reproductive and respiratory syndrome attenuated vaccine; Realmso-bidi-font-family: 宋体" lang="EN-US">-time PCR; application

MA Zhi-liang, SUN Ting-ting, YANG Jie, FAN Bin, LIU Xu-chuan, ZHANG Yi-fang. Development and Preliminary Application of Real-time PCR Assay for Detecting Quantitation of PRRSV in Vaccine[J]. .

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Development and Preliminary Application of Real-time PCR Assay for Detecting Quantitation of PRRSV in Vaccine

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