应用逆转录酶链反应对我国流行性出血热病毒分型

Chines Epidemic Hemorrhagic Fever Virus were Typed by RT-PCR Method

针对我国流行性出血热病毒的S片段核蛋白 (NP)编码基因保守核苷酸序列 ,合成了一对引物 ,扩增出编码核蛋白的 5 90bp碱基。扩增的II型产物存在限制性内切酶PstI的酶切位点 ,而I型产物不存在。因此 ,用酶切扩增产物的方法达到病毒分型的目的。试验提取 8份鼠脑传代毒种 ,1份细胞培养物和 2 5份病人血清中的出血热病毒RNA ,同时做 6份正常人血清对照 ,经逆转录PCR酶切分型。试验结果同间接免疫荧光技术 (IFA)分型结果相一致 ,可特异地对我国流行性出血热病毒进行分型。

According to the conserved nucleotide sequences of the gene coding for the nucleoprotein (NP) of hemorrhagic fever with renal syndrome virus Segment, A pair of primers were designed for PCR. The PCR products were identified with 590 nucleotide in with type II virus has a Pst I site and type I virus has not. So the hemorrhagic fever with renal syndrome virus type can be differentiated by this method. This experiment was performed by extracting 8 virus from rat brain, extracting 1 virus from cell culture, extracting 25 virus from relevant patients' serum and contrasting with 6 serum from normal people. During RT-PCR and Pst I cutting analysis. The results show the same conclusion as the results derived by IFA analysis.hemorrhagic fever with renal syndrome virus Fever virus type can be specifically differentiated by this method.;