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RT-PCR结合限制性片段长度多态性分析对我国汉坦病毒进行基因分型的研究
Study on genotyping of Hantavirus isolates of China by RT PCR/RFLP
【摘要】 用电脑软件分析HFRSV 囊膜糖蛋白编码基因的M 节段,寻找不同型别间病毒株M 节段的限制性内切酶酶切位点的差异。结果发现,HTN 型76118 株和SEO 型R22 株M 节段1199~1497 间的酶切位点,可用于进行限制性内切酶基因多态性分析,以确定HFRSV 的型别。根据上述结果,合成一对引物,建立RT/PCR 方法,分别用Rsal、Taq 1 和HindIII3 种内切酶对扩增片段进行酶切分型。用此方法分析了从我国不同地区、不同宿主分离的毒株19 株,及国际标准毒株1 株。结果表明,9 株可定为HTN 型,8 株可定为SEO 型,3 株无法确定其型别。此20 个毒株曾用血清学方法分型,仅11 株分型成功。分型成功率RTPCR/RFLP法与血清学方法分别为85% 和55% ,前者比后者高30 % 。
【Abstract】 The sequences of M segment genes of the prototype viruses of HTN and SEO serotypes were analysed, which indicated that there were different restriction endonucleas sites in sequence 1199~1497 which could be used for typing The present study reported that 20 Hantanvirus strains were typed by reverse transcriptional polymerase chain reaction(RT PCR) and restriction fragment length polymorphism(RFLP)analysis The results showed that of the 20 Hantanvirus strains, 9 and 8 strains were HTN and SEO, respectively, and the other 3 strains were unable to be determined The experiment suggested that the RT PCR/RFLP could be used for genomic typing in Hantanvirus successfully, while only 11 out of the 20 isolates could be typed by serological methods previously, and which showed that the typing rate by RT-PCR/RFLP and serological method were 85%and 55%, respectively, and the typing rate of the former was higher than that of the latter by 30%
- 【文献出处】 细胞与分子免疫学杂志 ,JOURNAL OF CELLULAR AND MOLECULAR IMMUNOLOGY , 编辑部邮箱 ,1999年04期
- 【分类号】Q789
- 【被引频次】5
- 【下载频次】62