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根据HFRSV汉滩型（HTN）代表株76－118和汉城型（SEO）代表株R22基因资料，设计了两组引物，用电脑软件分析证明设计符合引物标准。以一组引物克隆全长S基因片段和N端的部分S基因片段，并使它们在T7系统进行融合表达和非融合表达。非融合表达产量虽不及融合表达高，但生物活性好。以非融合表达的两个S基因片段产物作间接ELISA的包被抗原，其工作浓度均达1：100000用另一组引物建立了逆转录-聚合酶链式反应（RT-PCR），检测我国不同地区由8种主要宿主分离的37个HFRSV毒株，2个阳性标准对照毒株和5个阴性对照标本，并与cELISA法比较，二者阳性检出率分别为100％和84．6％，符合率为84．6％，但前者比后者敏感性高15．4％。对其中20个毒株的PCR扩增产物先后用RsaⅠ和HindⅢ作二级酶切，建立了逆转录-聚合酶链式反应-限制性片段长度多态性分析（RT-PCR－RFLP）分型法。被定为HTN型的9株，SEO型的8株，余3株未能定型。此20个毒株曾用血清学方法分型，仅11株分型成功．与RT-PCR－RFLP法结果符合。分型成功率RT-PCR－RFLP法比血清法高30％。
Two sets of primers were designed according to the data of HFRSV Hantaan (HTN) strain 76-118 and SEO representative strain R22, and the software was used to prove that the design conformed to the standard of primer. A set of primers was used to clone the full-length S gene fragment and the N-terminal partial S gene fragment, and to make them fused and non-fused in the T7 system. Although the yield of non-fusion expression is lower than that of fusion expression, its biological activity is good. The non-fusion expression of the two S gene fragment products as indirect ELISA coated antigen, the working concentration of 1: 100000 with another set of primers established by reverse transcription-polymerase chain reaction (RT-PCR), detection of our country 37 HFRSV isolates, 2 positive standard control strains and 5 negative control isolates from 8 major hosts in different regions of China and compared with the cELISA method, the positive rates of the two strains were 100% and 84.6% , The coincidence rate was 84.6%, but the former was 15.4% more sensitive than the latter. The PCR products of 20 strains were digested with RsaⅠand HindⅢ, and RT-PCR-RFLP was used to establish the reverse transcription-polymerase chain reaction-restriction fragment length polymorphism (RT-PCR-RFLP) law. Nine strains were classified as HTN type, eight as SEO type, and the remaining three strains failed to shape. The 20 strains have been serologically typed, only 11 strains were successfully typed. In accordance with the results of RT-PCR-RFLP method. The success rate of typing RT-PCR-RFLP method is 30% higher than that of serum method.