逆转录-套式PCR方法鉴定福建省登革热分离株型别的研究

Application of reversed-nest PCR to identify the sero-type of Dengue virus isolated from Fujian province

目的 应用分子生物学手段鉴别福建省登革热分离株的型别。方法 病毒培养上清经浓缩后离心沉淀 ,提取病毒ＲＮＡ ,行ＲＴ -ＰＣＲ ,再用型特异性引物扩增出特异性片段 ,电泳观察判断其型别。结果 从早期病人血液中分离的 3株和从白纹伊蚊中分离的 1株病毒经扩增后其产物均为登革热 2型特异性条带。结论 福建省 4株分离株均为登革热 2型病毒。

Aim Typing the isolates of Dengue virus from Fujian in China Method The RNA extracted from suspension of culture,was used to synthesize cDNA in a reversed transcriptase polymerase reaction(RT PCR) with a pair of universal primers To identify the sero type of isolates by the sizes of products yielded in a second round PCR amplification with type specific primers Result The products from 4 isolates were all the specific fragments of Dengue sero type 2 virus,including 3 from patient and 1 from mosquito,respectively Conclusion The 4 isolates of dengue virus from Fujian province are all identified to be Dengue sero type 2 virus