5种外来动物疫病并联荧光定量PCR检测方法的研究

摘要

According to the gene sequences of related viruses reported in Genbank,primers and probes were designed based on the conservative region of Peste des petits ruminants virus(PPRV)African swine fever virus(ASFV), West Nile fever virus(WNV),Hendra virus(HeV)and Nipah virus(NiV)(the conservative regions were N gene of PPRV,p72 gene of ASFV,H gene of HEV and NIV,and PRM gene of WNV,respectively). After screening a set of specific primers and a TaqMan probe,a real-time fluorescene quantitative PCR for detection of the above five exotic animal diseases was established. The clinical detection results of 103 samples used this method showed that all the samples were tested negative except the positive control,which verified the parallel fluorescene quantitative PCR of Taqman mode established in this study has characterisitics of differential diagnosis. The method can be used as a reference method for clinical detection of above-mentioned five kinds of pathogens.%根据 GenBank中发表的相关病毒的基因序列，通过分析比较小反刍兽疫病毒（PPRV）、非洲猪瘟病毒（ASFV）、西尼罗河热病毒（WNV）、亨德拉病毒（HeV）和尼帕病毒（NiV）的保守区域（PPRV的N基因、ASFV的P72基因、HeV和NiV的H基因，以及WNV的PrM基因），各设计筛选出一套特异性引物和Taqman探针，建立了5种外来动物疫病并联荧光定量RT-PCR检测方法。利用该方法，对103份样本进行了临床检测，结果除阳性对照外，其他均为阴性。检测结果验证了本研究建立的Taqman模式并联荧光定量PCR检测方法具有鉴别诊断的特点，可以作为临床检测这5种病原体的参考方法。