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目的：建立可用于汉坦病毒检测的ＲＴ－ＰＣＲ方法，并对其临床应用进行初步研究．方法：比较７６－１１８株和ＳＲ－１１株Ｓ基因序列，选择其保守区设计半套式引物，通过病毒培养上清及患者血清的扩增检测进行ＰＣＲ方法的敏感性、特异性及临床应用研究．结果：所建立的ＲＴ－ＰＣＲ方法可检出１．２５ＰＦＵ的病毒量；对照组（正常细胞培养上清和正常人血清）经扩增检测均为阴性；８６份经ＩＦＡ检测抗ＨＶ－ＩｇＭ阳性的肾综合征出血热（ＨＦＲＳ）患者血清，其阳性检出率按发病时间分别为：１ｗｋ内１００％；１ｗｋ～２ｗｋ６１．２９％；２ｗｋ～３ｗｋ３．３％；＞３ｗｋ为２８．５％．另外２０份ＩＦＡ（－）但疑诊ＨＦＲＳ的初入院患者血清的检出率为３０％，经进一步跟踪观察，确认为ＨＦＲＳ．结论：ＲＴ－ＰＣＲ是一种敏感、特异的ＨＦＲＳ诊断方法，可弥补血清学检测的不足，值得进一步完善、推广和应用．
Objective: To establish a RT-PCR method for detection of Hantavirus and to study its clinical application. Methods: The S gene sequences of 76-118 and SR-11 strains were compared. Semi-set primers were designed according to their conserved regions. Sensitivity, specificity and clinical application of PCR were determined by amplification of virus culture supernatant and serum of patients the study. Results: The established RT-PCR method could detect the virus quantity of 1.25 PFU; the control group (normal cell culture supernatant and normal human serum) were negative by amplification; 86 IFA-positive anti-HV-IgM The positive rate of hemorrhagic fever with renal syndrome (HFRS) was 100% in 1wk, 61.29% in 1wk ~ 2wk, 3.3% in 2wk ~ 3wk, and 28.5% in> 3wk. Another 20 IFA (-) but suspected HFRS patients admitted to the hospital early detection rate of 30%, after further follow-up observation, was identified as HFRS. Conclusion: RT-PCR is a sensitive and specific method for the diagnosis of HFRS, which can make up for the lack of serological detection and is worth further improvement, promotion and application.