河南省2008～2009年麻疹野病毒核蛋白基因特征分析

Analysis of the Nucleoprotein Gene on Measles Viruses Isolated in Henan Province in 2008-2009

目的了解河南省流行的麻疹野病毒的基因特征,为制定消除麻疹策略提供依据。方法用非洲绿猴肾细胞/淋巴信号激活因子转染的非洲绿猴肾细胞[Vero Cell/Signaling Lymphocyte Activation Molecule(Vero/SLAM)]从疑似麻疹病例标本中分离麻疹野病毒,用逆转录-聚合酶链反应(Reverse Transcription-Polymerase ChainReaction,RT-PCR)扩增麻疹野病毒分离株的核蛋白(Nucleoprotein,N)基因羧基末端676个核苷酸片段,再对扩增产物进行核苷酸序列测定和分析。以N基因羧基末端450个核苷酸片段构建基因亲缘性关系树,进行核苷酸变异分析。结果河南省2008～2009年共分离麻疹病毒147株,挑选其中73株测序后均为H1基因型,73株麻疹野病毒450个核苷酸的平均进化距离为0.020。2008年分离株与参考毒株的平均进化距离为0.055,其中与Hunan.China93-7/H1基因型的进化距离为0.012～0.026;2009年分离株与参考毒株的平均进化距离为0.067,其中与H1基因型的进化距离为0.013～0.020。结论河南省2008～2009年麻疹野病毒流行优势株为H1基因型,未发现病毒有较大变异,但存在一定的进化距离,呈现多个传播链的流行。

Objective To know the genic characteristics of wild type measles viruses in Henan province from 2008 to 2009,and to provide the evidence for measles control and eradication.Methods Measles viruses were isolated from the specimens collected from suspected measles cases by vero cell/signaling lymphocyte activation mdecule（Vero/SLAM）cell.Fragment of 676 nucleotide acids of the carboxyl end of nucleoprotein gene were amplified by reverse transcription-polymerase chain reaction（RT-PCR）,the PCR products were sequenced and analyzed.based on 450 nucleotide acids of the carboxyl end of nucleoprotein gene phylogenetic tree was constructed.Resuits 147 wild type measles viruses were isolated in Henan province from 2008 to 2009.73 strains were selected and identified as H1 genotype,the overall distance was 0.020.In 2008,the distance between the strains and the reference strains was 0.055.the distance between the strains and Hunan.China 93.7/H1 were 0.012-0.026.In 2009,the distance between the strains and the reference strains was 0.067,the distance between the strains and Hunan China 93-7/H1 were 0.013-0.020.Conclusions H1 genotype was predominant genotype of wild measles viruses circulated in Henan province from 2008 to 2009.Viruses did not have variation,but there was change of distance,which had several transsmission chain.