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目的 对分离自湖北省不同地区的 30株汉坦病毒 (HV)RNAM片段进行分析 ,以了解湖北省不同地区HV的基因特征及基因组功能。方法 参考HV的基因文库软件设计M片段G1 区型特异性引物 ,用适合于湖北省HV分型的逆转录 -半巢式聚合酶链反应 (RT -heminestedPCR) ,并结合限制性内切酶酶切图谱分析 ,对湖北省不同地区分离的 30株HV进行分型、酶切。结果 分离于湖北省不同地区的 30株HV的分型结果为汉滩型 2 1株、汉城型 9株。 2 1株汉滩型靶基因被HinfⅠ、HaeⅢ消化的图形与标准株 76/ 1 1 8株的图形一致 ,未见与H - 1 1 4株相似的图形 ,其中武汉和洪湖各有l株靶基因HaeⅢ消化只见 1个片段 ;应城、江夏、黄陂和麻城各有 1株靶基因HinfⅠ消化有 3个片段。另外 2 1株汉滩型靶基因除前述的应城毒株外均被HincⅡ消化。 9株汉城型靶基因不能被HaeⅢ消化 ;HinfⅠ消化只见 2个片段 ,而EcoRⅠ消化却见 2个片段 ,与R2 2 株消化的图形不一致。结论 湖北省HFRS流行为混合疫区 ;汉城型M片段基因稳定 ,汉滩型M片段基因不稳定 ,且在湖北地区具有一定的地理聚族性。
Objective To analyze the HVM RNAM fragments of 30 isolates from different regions of Hubei Province to understand the genetic characteristics and genomic functions of HV in different regions of Hubei Province. Methods The genomic DNA library of M segment was designed by reference to the HV gene library software. The RT-PCR was performed with RT-heminested PCR, which was suitable for HV genotyping in Hubei province. Combined with the restriction endonuclease Cut map analysis, the separation of 30 different regions in Hubei Province HV typing, digestion. Results The genotyping results of 30 HV isolates isolated from different regions of Hubei Province were 21 strains of Han Tan and 9 strains of Seoul. The pattern of Hantaan - type target gene digested by Hinf Ⅰ and Hae Ⅲ was the same as that of the standard strain 76/118, but there was no similar pattern to that of H - 114 strain, in which Wuhan and Honghu each had 1 target One fragment was found in the digestion of Hae Ⅲ gene and one fragment of HinfⅠ gene in Yingcheng, Jiangxia, Huangcheng and Macheng respectively. In addition, 21 strains of Hantaan target genes were digested by Hinc Ⅱ except the above-mentioned strain Yingcheng. Nine strains of Seoul-type target genes could not be digested with Hae Ⅲ; two fragments were found by HinfⅠ digestion and two fragments by EcoRI digestion, which were inconsistent with the digestion patterns of R2 2 strains. Conclusions HFRS prevalence in Hubei Province is mixed epidemic area. The gene of Seoul M gene is stable, and the gene of Hantaan M gene is unstable, and it has some geographical polygamy in Hubei Province.