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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 Hantavirus infection early. Methods According to the international standard strains (76 118, 80 39) M gene G1 region designed two pairs of public primers and a probe, according to the one-step extraction of hantavirus isocyanurate-phenol, RNA detection using RT nested PCR virus Cell culture supernatants and dilutions, 16 control serum and 40 hantavirus RAN in 40 acute HFRS serum samples (1 to 5 days). All amplified products were confirmed by Southern blot or Dot blot. Results The specific fragment of 453bp was amplified from the Hantaan virus cell culture medium and 38 of the 40 acute HFRS serum samples, while 10 normal serum samples and 12 non-HFRS serum samples were negative. The method sensitivity Higher than cell culture. Conclusion RT-nested PCR is a sensitive, specific and rapid method for the early diagnosis of Hantavirus infection.