基于实时荧光定量PCR技术的猪伪狂犬病活疫苗含毒量评价

doi:10.16431/j.cnki.1671-7236.2019.04.021

Abstract

Abstract:

In order to quantitatively analyze the virus content of clinical live pseudorabies (PR) vaccine in Guizhou province,a pair of specific primers was designed according to the sequence of pseudorabies virus (PRV) gB gene in GenBank (accession No.:M17321.1),a Real-time quantitative PCR method was established,and the virus content of live PR vaccine from 6 different manufacturers was analyzed.The results showed that a Real-time quantitative PCR method was successfully established for detection of PRV.There was a good linear relationship between the number of plasmid copies diluted in the standard curve and Ct value,the standard curve equation was Y=-0.97X+33.66 (R²=0.999).The method had a minimum detection virus content of 3.19×10¹ copies/μL,high sensitivity,good repeatability and specificity.The Real-time quantitative PCR method was applied to detect the viral content of live PR vaccine produced by 6 manufacturers in Guizhou province.The results showed that the virus content of 6 live PR vaccine (A-F) sold in 6 markets were 1.67×10⁷,4.83×10⁵,2.64×10⁶,4.27×10⁷,3.39×10⁶ and 3.68×10⁵ copies/μL,the viral content of the vaccine from different manufacturers was significantly different,the biggest difference could be 116 times.Two kinds of live PR vaccine from A and D manufacturers had higher viral content.The Real-time quantitative PCR method established in this experiment could be used for rapid detection and evaluation of the viral content of live vaccine against porcine PR,it had a certain guiding significance for the quality control of the live vaccine of PR and the selection of clinical vaccine.

Key words: pseudorabies (PR); vaccine; virus content; evaluation

CLC Number:

S852.65⁺9.1

WANG Bolin, ZHOU Yi, HE Ling, YANG Mei, CHENG Zhentao. Evaluation of Virus Content of Live Pseudorabies Vaccine Based on Real-time Quantitative PCR[J]. , 2019, 46(4): 1135-1142.

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Evaluation of Virus Content of Live Pseudorabies Vaccine Based on Real-time Quantitative PCR

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