猪圆环病毒多重PCR检测与分型

Multiplex PCR Detection and Typing of Porcine Circovirus

[目的]建立一种快速的PCV诊断方法,并能将2种PCV病毒区分开来,提供兽医临床和相关产品快速监测PCV的手段。[方法]根据genebank上公布的PCV1和PCV2序列,设计2对引物扩增PCV1和PCV2非同源性区域,建立快速检测PCV2个血清型的多重PCR,用已知的PCV1和PCV2及其他病毒细菌检测其敏感性和特异性。[结果]结果表明:所建立的方法特异、敏感,最低能检出0.02 ng的PCV1和PCV2混合DNA,用该法检测已知阳性病料和不同来源的猪血清,阳性符合率达100%。[结论]所建立的MultiplexPCR可用于PCV2个血清型的临床快速检测。

The objective of this experiment is to detect and type two serovars of PCV with multiplex polymerase chain reaction （Multiplex PCR）, two sets of primers were designed following the comparison of DNA sequences of porcine eireovirus （PCV） types 1 and type 2 from the Gene bank. The reaction conditions of Multiplex PCR was optimization. The sensitivity and the specificity were tested using swine viruses and bacterium, in addition to PCV1 and PCV2. The results showed that the method is sensitive and specific, as low as 0.02 ng total DNA of PCV1 and PCV2 could be measured. The Multiplex PCR method was also used to detect swine sera with PMWS and non- PMWS form Hainan, Jiansu and Shixing of Guangdong. The accuracy for 4 positive sera from Jiangsu was 100 %.The result suggested that the Multiplex PCR assay can be used to rapidly detect and type PCV for clinical veterinary medicine.