Detection Of PRV,PPV And PCV2 Infection Using PCR

A multiplex ploymerase chain reaction was optimized to simultaneousely detecte three pathogens of PCV2,PPV and PRV.Three sets of specific primers were designed according to the conservative sequences of PCV2,PPV and PRV.It was proved that the samples which contained PCV2,PPV and PRV could be amplified by the multiplex PCR using the three sets primers when MgCl₂,TaqDNA polymerase and dNTP density were optimized separately.Double ploymerase chain reaction was carried on,in the identical sample random sets primers(PCV2 and PPV,PCV2 and PRV,or PPV and PRV)can be amplified.In the last multiplex ploymerase chain reaction was optimized,In the identical sample it can be simultaneously amplified PCV2,PPV and PRV' specific primers.it would yield three specific bands of 950bp(PCV2),490bp(PPV)and 372bp(PRV).But no specific bands were amplified from other pathogenic viruses and bacterium.As little as PCV2 1.4pg/l,å’ŒPPV 0.22pg/lå’ŒPRV 34.7pg/l.were detected using gelelectrophoresis in this multiplex PCR..This method could differentiate PCV2 infection,PPV infection,PRV infection.and mixed infection from the reproductive failure.Since October,2006 in 15 intensified pig farms of Henan and Shandong,75 samples were examied.Lung,liver,spleen,lymph node and blood serum can be examied DNA of PCV2.The masculine rate was between 33.3-100%.The masculine rate of all samples was 61.7%。Clinical symptom of pigs was diverse.75%of examined pigs became emaciated,the growth of pigs was slow,appetite of pigs droped,the spirit of pigs missed.50%of examined pigs had breath symptom.Some pigs had anemia and jaundice.A few had diarrhea.One of examined pig farm in Rizhao had typical PMWS symptom.The masculine rate is 100%。Organ of 2 days pigs in Luoyang can examine PCV2.Blood serum of sick pigs in Nanyang can examine PCV2.sick pigs had not Clinical symptoms.