摘要
猪瘟病毒(CSFV)、流行性乙型脑炎病毒(JEV)和猪繁殖与呼吸综合征病毒(PRRSV)是引起严重的种猪繁殖障碍的病原,而且经常混合感染,及时准确诊断是防治的前提。根据GenBank发表序列选取3对引物建立检测CSFV、JEV和PRRSV病毒的多重RT-PCR方法,扩增产物分别为508 bp、380 bp、263 bp。经与IDEXX商品化的检测CSW抗原试剂盒比较,二者的符合率为96.7%;扩增JEV和PRRSV PCR产物分别经EcoR V和Sau3A I酶切得到预期的片段。建立的多重RT-PCR检测JEV、PRRSV和CSFV敏感度分别为12.5个TCID_(50)、10个TCID_(50)和10^(-3)ng总RNA。结果表明该多重RT-PCR方法具有很好的特异性和敏感性,可用于临床三种病毒核酸的检测。
Classical swine fever virus （CSFV）, Japanese encephalitis virus （JEV） and porcine reproductive and respiratory syndrome virus （PRRSV） are the major causes of some serious reproductive diseases and co-infection is common. Therefore rapid and reliable diagnosis is crucial for effective prevention and treatment. In this study, a multiplex PCR was developed using primers designed according to published CSFV, JEV and PRRSV sequences. PCR products of 508 bp, 380 bp and 263 bp were obtained for respective viruses. Restriction digestion of PCR products with EcoR Ⅴ and Sau3A Ⅰ produced expected target DNA fragments. Compared to CSFV antigen test kit, the multiplex RT-PCR showed 96.7 % coincidence with the commercial kit based on clinical test results. As low as 12.5 TCID50 JEV, 10 TCID50 PRRSV and 10^-3 ng total RNA including CSFV RNA could be detected by multiplex PCR. The result showed that the Multiplex RT-PCR is a rapid, sensitive and specific method for detecting virus from clinical samples.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2007年第5期385-388,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家863项目(编号:2001AA249012)
国家攻关项目(编号:2004BA514A16-5)资助
关键词
猪瘟病毒
流行性乙型脑类病毒
猪繁殖与呼吸综合征病毒
多重RT-PCR
classical swine fever virus
Japanese encephalitis virus
porcine reproductive and respiratory syndrome virus
multiplex RT-PCR
