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建立准确、快速、灵敏的汉坦病毒基因分型方法，可弥补传统血清学方法的不足，对防治该病毒所致的肾综合征出血热(HFRS)具有重要意义。本试验采用逆转录－聚合酶反应(RT-PCR)和限制性片段长度多态性(RFLP)和限制性片段长度多态性(RFLP)方法，对流行于我国的两型汉坦病毒代表株??汉滩型(HTNV)76－118和汉城型(SEOV)R22株进行基因分析。根据病毒DNA序列的电脑软件分析，不同HFRSV囊膜糖蛋白编码基因M节段1199~1497间核苷酸序列上RsaI、TaqI和HindIII的酶切位点存在差异(Fig.1), 可用于进行限制性内切酶基因多态性分析，以确定HFRV的型别。首先，以一对引物扩增该片段(Fig.2)。然后，分别用这三种内切酶(Fig.3,4,5)进行酶切分型。共分析了从我国不同地区，不同宿主分离的毒株18株，及国际标准毒株2株，酶切图谱显示，这些毒株可以被分为三组(Table.1)：9株可定为HTNV型，8株可定为SEOV型，3株无法确定其型别(X型)。该法分型结果与血清学经典的空斑减数中和试验分型结果基本一致，说明该酶切分型方法具有一定的可行性。
The establishment of an accurate, rapid and sensitive genotyping method of Hantavirus can make up for the deficiency of traditional serological methods and is of great significance for the prevention and treatment of hemorrhagic fever with renal syndrome (HFRS) caused by this virus. In this study, RT-PCR, RFLP and RFLP were used to detect the expression of two types of Hantavirus strains Han-type (HTNV) 76-118 and Seoul-type (SEOV) R22 strains were genotyped. According to the computer software of the viral DNA sequence analysis, the restriction sites of RsaI, TaqI and HindIII on the nucleotide sequence of 1199 to 1497 in M segment of HFRSV envelope glycoprotein were different (Fig.1) Restriction enzyme gene polymorphism analysis to determine the type of HFRV. First, the fragment is amplified with a pair of primers (Fig. 2). Then, the three kinds of endonucleases (Fig.3,4,5) were used for restriction enzyme digestion. A total of 18 strains of isolates isolated from different hosts in China and 2 isolates of international standard were analyzed. The enzyme digestion patterns showed that these isolates could be divided into three groups (Table 1): 9 strains could be defined as HTNV type, 8 strains can be classified as SEOV type, 3 strains can not determine the type (type X). The result of this method was basically the same as that of serological classic plaque meosis neutralization test, which indicated that the method of digestion and typing was feasible.