肾综合征出血热病毒逆转录－聚合酶链反应检测方法的实验研究和临床意义  

Experimental Study on Reverse Transcription-Polymerase Chain Reaction for Detection of Hemorrhagic Fever with Renal Syndrome virus and its Clinical Significance

作　　者：辛宏[1] 于洋[1] 蔡龙荣[1] 何建文[1] 叶芳耘 

机构地区：[1]上海第二军医大学长海医院临床实验诊断科

出　　处：《中华实验和临床病毒学杂志》1995年第4期341-343,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：上海市科学技术发展基金

摘　　要：选择覆盖肾综合征出血热病毒（ＨＦＲＳＶ）核蛋白（ＮＰ）基因的保守区核苷酸序列合成引物，采用异硫氰酸胍一步法抽提ＲＮＡ，建立了逆转录（ＲＴ）聚合酶链反应（ＰＣＲ）检测临床血清样本中ＨＦＲＳＶＲＮＡ的方法。扩增产物经凝胶电泳及Ｓｏｕｔｈｅｒｎ印迹杂交证实了其具有特异性。结果显示ＨＦＲＳＶ阳性细胞培养液、１６份患者血清均为阳性，而１０份正常人血清均为阴性。该方法具高特异、高敏感的特点，为临床早期、快速诊断，并从分子生物学角度鉴定ＨＦＲＳＶ提供了新手段。Hemorrhagic fever with renal syndrome virus(HFRSV) has a tripartite, single-stranded, negative-sense RNA genome.Two 20-mer oligonucleotides flanking the s segment were chosen as primers. RNA, extracted by single-step method of acid guanidinium thiocyanate-phenol-chloroform; A reverse transcription-polymerase chain reaction(RT-PCR) was adopted for detection of HFRSV in viral infected Vero-E6 cells and in clinical serum speimens. The PCR product was then detected by agarose gal electrophoresis and southern hybridization. Serum samples from all the sixteen patients showed positive result.while the normal serum samples showed negtive result. It was suggested that the detection method of RT-PCR is simple, sensitive, specific and can be applied for clinical detection.

关 键 词：肾综合征出血热 聚合酶链反应 RNA 出血热病毒 

分 类 号：R512.804[医药卫生—内科学]