中国兽医科学2011，Vol.41Issue(3)：267-272,6.
猪繁殖与呼吸综合征病毒变异株实时荧光定量RT-PCR检测方法的建立
Development of real-time fluorescent quantitative RT-PCR assay for detection of porcine reproductive and respiratory syndrome virus variant strains
高诗敏 1王敏功 1邓瑞林 2姚晓辉 2王林川1
作者信息
- 1. 华南农业大学,广东,广州,510642
- 2. 广东温氏集团,广东,云浮,527439
- 折叠
摘要
Abstract
To identify highly pathogenic porcine reproductive and respiratory syndrome virus(PRRSV) variants rapidly using real-time PCR technique, primers and TaqMan probe for specific PCR amplification were designed based on sequence differences between variant strains and classical strains.The fluorescent quantitative RT-PCR assay for detection of PRRSV variants was established.Its specificity,sensitivity and stability were examined.This assay had differentiate PRRSV variants.1 TCID50 of template RNA could be detected.The sensitivity of this assay was 100 times higher than the conventional RT-PCR.Content could be absolutely quantified of virus.The variation coefficients were less than 10％ based on 4 replications of each sample.13 lung samples were examined by the developed fluorescent quantitative RT-PCR and the conventional PCR, respectively,and 11 were detected to be infected with PRRSV by the real-time PCR, but only 9 were confirmed to be infected with PRRSV by the conventional PCR.These results showed that the developed fluorescent quantitative RT-PCR assay was rapid, specific, sensitive and simple for the detection of PRRSV variants and the identification of highly pathogenic PRRSV.关键词
猪繁殖与呼吸综合征病毒/变异株/实时荧光定量RT-PCR分类
农业科技引用本文复制引用
高诗敏,王敏功,邓瑞林,姚晓辉,王林川..猪繁殖与呼吸综合征病毒变异株实时荧光定量RT-PCR检测方法的建立[J].中国兽医科学,2011,41(3):267-272,6.
