猪繁殖与呼吸障碍综合征病毒(PRRSV)检测方法的建立  

The establishment of detecting method for porcine reproductive and respiratory syndrome virus(PRRSV)

作　　者：苏晓鸥[1] 李玉荣[2] 乔俊文[1] 赵德明[1] 

机构地区：[1]中国农业大学动物医学院国家动物海绵状脑病实验室,北京100193 [2]河北农业大学动物科技学院,河北保定071000

出　　处：《河北农业大学学报》2009年第1期82-85,共4页Journal of Hebei Agricultural University

基　　金：科技部:兽医微生物菌种资源标准化-朊病毒类资源标准化整理整合及共享(2005KDA21205-7);实验用小型猪地方标准研究;北京市科委(D07080200720701)

摘　　要：根据GeneBank中已经发表的PRRSV毒株的ORF6基因高度保守序列，设计并合成了一对特异性引物，建立了用于检测PRRSV的RT—PCR诊断方法。从感染PRRSV—QD的Marc-145细胞中提取病毒RNA，然后经RT—PCR扩增，获得预期266bp的目的片断，而猪伪狂犬病毒（PRV）、猪细小病毒（PPV）、猪圆环病毒（PCV）细胞培养物及Marc-145细胞进行同条件检测，结果为阴性；PRRSV扩增产物测序结果与GeneBank中已发表的PRRSV毒株序列同源性达94．4％～97．4％且应用本研究检测方法对已知基因型的10株PRRSV盲检，检出率100％。上述研究结果表明，所建立的RT—PCR诊断方法特异性好、敏感性高，可进一步应用于猪繁殖与呼吸综合征的临床诊断。Based on highly conservative ORF6 gene sequence of PRRSV on GeneBank, a pair of primers was designed and synthesized. The RT- PCR method for detecting PRRSV was established. PRRSV- QD RNA was extracted from virus - infected Marc - 145 cell and then amplified by RT- PCR. The positive 266bp specific fragment was obtained as expected. Under the same conditions, however, the results for detection of PRV, PPV, PCV and Marc - 145 cell by RT - PCR were all negative. The amplified sequence of PCR product of PRRSV - QD shares 94.4 % - 97.4 % homology with the published sequence of other PRRSV strains from GeneBank. The results indicate that RT- PCR method in this research has high sensitivity and specificity, and could be further applied for PRRSV clinical diagnosis.

关 键 词：猪繁殖与呼吸综合征 PRRSV ORF6 RT-PCR 检测 

分 类 号：S852.65[农业科学—基础兽医学]