猪血凝性脑脊髓炎病毒RT-PCR方法的建立及初步应用

Establishment of Detection of Hemagglutinating Encephalomyelitis Virus by RT-PCR

为了建立快速检测猪血凝性脑脊髓炎病毒(HEV)的RT-PCR方法,根据GenBank中HEV的HE基因和S基因设计了1对引物,在优化RT-PCR反应条件的基础上,成功的扩增出323bp的特异性条带。检测与HEV亲缘性较高的牛冠状病毒及猪的伪狂犬病毒均为阴性,最低可以检测到10个TCID50/100μl的病毒,说明该方法的有较好的特异性和敏感性。用此RT-PCR方法对感染HEV的小鼠和猪进行检测,结果能从发病动物的多种组织中检测到病原,其中以脑组织的检出率最高。因此,临床疑似病例检测时以脑组织为最佳检测样本。

In order to establish the detection of Hemagglutinating Encephalomyelitis Virus （HEV） by RT- PCR, a pair of oligonucleotide primers appropriate for PCR amplication was selected based on the HE gene and S gene of HEV. A reverse transcription - polymerase chain reaction （RT-PCR） assay was developed for the detection of HEV after optimization of the reaction conditions. The RT-PCR was sensitive and specific for HEV and did not amplify closely related bovine coronavirus and pseudorabies virus of pigs. The HEV can be found in many organs, especially found regularly in brain. So the brain of suspected pig is the best sample to detect.