多重PCR同时检测猪圆环病毒2型,猪细小病毒,猪繁殖与呼吸综合征病毒和猪瘟病毒

本试验建立一种可同时检测猪圆环病毒2型(PCV-2),猪细小病毒(PPV),猪繁殖与呼吸综合征病毒(PRRSV),猪瘟病毒(CSFV)4种病毒的多重PCR方法.对于每一种特定的病毒,用4对寡核苷酸引物均能特异扩增其目的片段.以含有病毒目的片段的质粒为模板,测定了多重PCR的检测灵敏度,PRRSV和CSFV检测最低限是48 pg,而PPV和PCV-2为0.48 pg.利用建立的多重PCR方法对具有产自有繁殖障碍母猪的仔猪或具有呼吸障碍症状的76个仔猪样本进行检测.检出了4种病毒的存在,其中26个样本(34.2%)同时感染了2种以上病毒.结果表明多重PCR方法检测猪混合感染的病毒,是一种快速、灵敏、低成本、高效率的病原学诊断工具.

A multiplex PCR for simultaneous detection of porcine circovirus type 2, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus in pig

A multiplex polymerase chain reaction(PCR)-based method was designed for simultaneous detection of four main viruses involved in respiratory and/or reproductive failure in piglets(porcine circovirus type 2 ,porcine parvovirus ,porcine reproductive and respiratory syndrome virus ,and classical swine fever virus ).Each of the four pairs of oligonucleotide primers exclusively amplified the targeted fragment of the specific viruses.The sensitivity of the multiplex PCR using purified plasmid constructs containing the specific viral target fragments was 48 pg for PRRSV and CSFV,0.48 pg for PPV and PCV-2.A total of 76 specimens from piglets with symptoms of respiratory and/or reproductive failure were collected in China during June 2006 to July 2007.All specimens were detected at least one of 4 viruses by multiplex PCR.Coinfections with these viruses were identified in 26 specimens(34.2%).Therefore,this method may be a rapid,sensitive,and effective etiological diagnostic tool for the routine surveillance of viral infections in pigs.