摘要
为建立同时检测猪瘟病毒(CSFV)和猪繁殖与呼吸综合征病毒(PRRSV)经典株和高致病性株(HP-PRRSV)的多重RT-PCR检测方法,本研究根据GenBank中登录的PRRSV经典株、HP-PRRSV株和CSFV基因组全序列,分别设计了2对特异性引物PRRSV-S/PRRSV-A和CSFV-S/CSFV-A,产物分别为544 bp、457 bp和294 bp。检测结果显示,该方法具有高特异性,均可检测到1 TCID50的病毒量。对65份临床上疑似为CSFV和PRRSV感染的病料样品进行检测的结果表明,本研究建立的多重RT-PCR检测方法可以用于临床上CSFV和PRRSV感染引起的传染病病原学的检测,并且能够准确鉴别PRRSV经典株及HP-PRRSV。
To establish the assay for detections of classical swine fever virus （CSFV） and porcine reproductive and respiratory syndrome virus （PRRSV）, as well as distinguishing the conventional PRRSV and the highly pathogenic PRRSV （HP-PRRSV）, the multiplex RT-PCR was developed with two pair specific primers of PRRSV-S/PRRSV-A and CSFV-S/CSFV-A designed for HP-PRRSV, conventional PRRSV and CSFV based on the sequences in GenBank. The results showed that the assay was able to detect CSFV, HP-PRRSV and conventional PRRSV simultaneously by amplifying DNA fragments of 294 bp, 457 bp and 547 bp, respectively, and the sensitivity of the assay was up to 1 TCIDs0 for each virus. Moreover, a total of 65 clinical samples tested by the multiplex RT-PCR, and the results were similar to the detections by commercial detection kits which the coincidence rate was 96.92%. Therefore, the multiplex RT-PCR assay could be used for detection of CSFV or PRRSV in infected pigs.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2014年第1期34-37,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
信阳农业高等专科学校青年教师科研基金项目(2009ZRKX016)
关键词
猪瘟
猪繁殖与呼吸综合征
多重RT—PCR
classical swine fever
porcine reproductive and respiration syndrome
multiplex RT-PCR
