安徽省蚌埠市麻疹野毒株核蛋白基因检测及序列分析

    Nucleoprotein gene detection and sequence analysis of wild-type measles strains from patients in Bengbu,Anhui

    • 摘要: 目的:通过检测麻疹野毒株的基因型,分析安徽省蚌埠市麻疹病毒流行株的基因特征,为麻疹监测和防治提供依据。方法:选取2010~2011年蚌埠市散发的15例临床确诊为麻疹的患者,收集患者急性传染期的尿液标本,提取病毒RNA,用RT-PCR方法扩增麻疹病毒核蛋白基因羧基末端的515 bp核苷酸片段,对扩增产物进行核苷酸序列测定和分析,构建基因亲缘关系树。结果:15份麻疹患者尿液标本中有14份分离到麻疹野毒株;将其中3株麻疹野毒株的核蛋白(N)羧基末端515 bp核苷酸与Genebank中麻疹病毒的8个参考株进行基因同源性关系比较,与Genebank中公布的H1、H2基因型参考株Hunan.CHN93-7、Beijing.CHN94-1所对应的核苷酸序列的遗传距离分别为0.020~0.026和0.064~0.075;而与H1a参考株China93-2的遗传距离最近,为0.009~0.015;3株麻疹野毒株N羧基末端515个核苷酸的同源性为97.8%~98.9%;与H1参考株Hunan.CHN93-7、H1a参考株China 93-2和H1b参考株China 94-7的核苷酸同源性分别为97.4%~98.0%、98.5%~99.1%和96.5%~97.1%。结论:蚌埠市麻疹野毒株的优势基因型为H1a基因亚型。

       

      Abstract: Objective: To provide basis for the monitoring and prevention of measles by detecting the genotype of wild-type measles strains and analyze the gene characteristics of epidemic measles virus from in Bengbu,Anhui.Methods: Fifteen sporadic cases clinically diagnosed as measles from 2010 to 2011 in Bengbu Anhui were selected in the study.The patients' urine samples were collected in the acute infectious period,the virus RNA was extracted,the nucleotide protein gene of carboxyl-terminal 515 bp nucleotide fragment of measles virus was amplified by RT-PCR,and gene phylogenetic tree was constructed by nucleotide sequencing and analysis of PCR.Results: Forteen wild-type measles strains were isolated from the urine samples of the 15 cases of measles;the gene homology of the 515 bp fragment of nucleoprotein(N) carboxy-terminal was compared between 3 wild-type measles strains and 8 reference strains in Genebank.The corresponding genetic distance of the nucleotide sequence to the published H1,H2 genotype reference strains in Hunan,CHN93-7,Beijing and CHN94 in Gene bank was 0.020 to 0.026 and 0.064 to 0.075 respectively;the distance to H1a reference strain China 93-2 was the shortest,which was 0.009 to 0.015;the 515 nucleotide homology of the N gene carboxy-terminal of the 3 measles strain were 97.8% to 98.9%;the corresponding nucleotide homology to the reference strains H1 CHN93-7,H1a China 93-2 and H1b China 94-7 were from 97.4% to 98.0%,98.5% to 99.1% and 96.5% to 97.1%,respectively.Conclusions: The dominant genotype of wild-type measles strains is H1a.

       

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