山东省部分地区肠道病毒71型的RT-PCR检测及基因特征分析  

RT-PCR diagnosis and analysis of the genetic characteristics of enterovirus 71 in parts of Shandong Province,China

作　　者：王世富[1] 张乐海[1] 马丽霞[1] 刘霞[1] 姜忠强[1] 于华凤[1] 

机构地区：[1]山东大学齐鲁儿童医院检验科,山东济南250000

出　　处：《中国病原生物学杂志》2009年第12期884-886,共3页Journal of Pathogen Biology

摘　　要：目的采用逆转录聚合酶链反应（RT-PCR）检测2008年山东省部分地区手足口病（HFMD）患者肠道病毒71型（EV71）,对毒株进行诊断及基因特征初步分析。方法以肠道病毒特异引物对EV71进行RT-PCR扩增,经琼脂糖凝胶电泳鉴定阳性标本,纯化后与pGEM-T载体连接,转化大肠埃希菌DH5α,筛选后测序,所得序列与我国阜阳、深圳、台湾株及美国EV71标准流行株的核苷酸序列在线进行BLAST比对,分析其同源性。结果42份标本中检出EV71 33例（78.57%）,所得序列经种系进化分析,与阜阳2008年HFMD暴发分离株同源性为98.7%99.1%,与深圳1998年HFMD散发分离的EV71毒株同源性为93.4%93.8%,与台湾1998年HFMD病例分离毒株同源性为91.2%91.6%,与美国EV71毒株同源性为79.2%79.7%。有9株EV71的563位苏氨酸（T）替代了EU703813的异亮氨酸（I）。结论EV71 C4亚型是2008年山东省部分地区HFMD暴发流行的病原,同源性分析表明山东省部分地区EV71分离株与阜阳、深圳及台湾地区分离株有较高的同源性,提示该病毒在中国大陆有较广泛的传播。Objective To test for enterovirus 71（EV71）,which caused hand,foot and mouth Disease（HFMD） in areas of Shandong Province,China in 2008,using reverse transcription polymerase chain reaction（RT-PCR） and to determine and analyze the strain＇s genetic characteristics.Methods EV71-specific primers were used as the primers for RT-PCR.Positive specimens were identified by agarose gel electrophoresis,and the products were purified and linked to the pGEM-T vector.The vector was transformed into Escherichia coli DH5α for screening and then sequenced.Via online BLAST,its sequence was compared to the standard nucleotide sequence of the EV71 strains prevalent in Fuyang and Shenzhen,Taiwan,and the United States.Results Of 42 specimens,EV71 was detected in 33（78.57%）.Compared to the strain from Fuyang in 2008 and the strains from Shenzhen and Taiwan in 1998,the homology of the sequence was 98.7%-99.1% versus 93.4%-93.8% and 91.2%-91.6%.In comparison to the strain from the United States,the homology of ...更多the sequence was only 79.2%-79.7%.The amino acid isoleucine（I） in EU703813 at site 563 was replaced by threonine（T） in all 9 strains.Conclusion The EV71 C4 subtype caused the HFMD outbreak in 2008 in some areas of Shandong Province.Homology analysis showed that EV71 from these areas of Shandong Province has a high level of homology with that from Fuyang,Shenzhen,and Taiwan,suggesting that the virus has spread widely on the Chinese mainland.

关 键 词：手足口病 肠道病毒71型 逆转录聚合酶链反应 基因特征 

分 类 号：R373.2[医药卫生—病原生物学]