摘要
目的 早期诊断汉坦病毒感染。方法 根据国际标准株(76118 ,8039) M 基因G1 区设计两对公有引物和一条探针,按异硫酸氰胍—酚一步法提取汉坦病毒RNA,采用RTnested PCR检测病毒细胞培养上清原液及稀释液、16 份对照组血清、40 份HFRS急性期(1 ~5 天) 血清标本中的汉坦病毒RAN,所有扩增产物采用Southern blot 或Dot blot 证实其特异性。结果 汉坦病毒细胞培养液和40 份HFRS急性期血清标本中的38 份均可扩增出特异性的片段453bp ,而10 份正常人血清和12 份非HFRS血清均为阴性,该方法敏感度高于细胞培养。结论 RTnested PCR 是一种敏感、特异、快速的方法,可用于汉坦病毒感染的早期诊断。
Objective To diagnose the infection of Hantaan virus early.Method According to the published sequence of the M gene G\-1 segments of Hantavirus strain 76 118 and Seoul virus strain 80 39,we designed two sets of genus sepcial primer and a probe.Hantaan virus RNA was extracted using a guanidine isothiocyanate procedure.RT nested PCR was used to amplify the special frament from Hantaan virus cell culture supernatant,16 negtive control samples,40 HFRS serum samples of acute stage(1 5d) patients.All amplified products were confirmed by Southern Blot or Dot Blot.Results The 453bp sequence of Hantaan virus M gene G\-1 was specially amplified from Hantaan virus cell culture supermatant.Hantaan virus RNA was also detected in 38 of 40 HFRS acute stage serum samples.No positive results were found by PCR while detecting 10 normal human serum samples and 12 non HFRS patients serum samples.The sensitivity of RT nested PCR in detecting Hantaan virus infection is superior to that of virus cell culture.Conclusion These results indicate that RT nested PCR is a special and sensitive method in diagnosing the infection of Hantaan virus early.(Shanghai Med J,1999,22:692 694)
出处
《上海医学》
CAS
CSCD
北大核心
1999年第11期692-694,共3页
Shanghai Medical Journal
关键词
汉坦病毒
聚合酶链反应
M基因
肾综合征出血热
Hantaan virus Polymerase chain reaction M gene Segments G\-1 Hemorrhage fever with renal syndrome