目的 了解安徽省某高校一起急性胃肠炎暴发疫情的病原体基因分型和分子特征。方法 采用实时荧光定量PCR法(Real-time PCR)对6份粪便/肛拭子标本检测诺如病毒核酸,检出的阳性标本经传统RT-PCR扩增衣壳蛋白区(ORF2)部分序列、基因测序和型别鉴定,利用ClustalX 2.0和Mega 6.0软件对测定序列进行核苷酸同源性比对和构建基因进化和亲缘性关系树。结果 检出4份标本诺如病毒核酸阳性,阳性率为66.67%(4/6);基因序列比对和进化亲缘性关系显示:诺如病毒基因型为GII.4型;2毒株序列之间核苷酸同源性为100%;与2014～2015年上海株、香港株核酸同源性最高,均为100%;与2012澳大利亚Sydney株(JX459908)同源性为99.6%;与上海和香港两地区毒株亲缘性关系最近,聚为独立一簇,同属GII.4/Sydney 2012分支。结论 引发该起急性胃肠炎暴发的病原体为GII.4型诺如病毒,且属于GII.4 Sydney 2012变异株。
Abstract
Objective To understand the genotype and molecular characteristics of the pathogens associated with the epidemic outbreak of acute gastroenteritis in a university of Anhui province. Methods A total of 6 fecal(anal swab)samples from acute gastroenteritis outbreak were collected. Norovirus (NoV) nucleic acid was detected by using Real-time PCR, and the partial ORF2 of the all positive specimens were amplified by RT-PCR and sequenced. Phylogenetic tree was constructed by using Mega6.0 based on partial ORF2 sequences of norovirus to perform phylogenetic analysis. Results Of the 4 specimens, 4(66.67%)were positive for NoV, and two positive samples were sequenced successfully. Phylogenetic analysis showed that 2 strains belonged to genotype GII.4/Sydney 2012 norovirus. The nucleotide homology between the 2 partial ORF2 sequences was 100%. The regions had 99.6% and 100% nucleotide amino identity compared with Sydney 2012 strain (JX459908) in Australia and Shanghai/HK strains in 2014-2105, respectively. Conclusion The epidemic outbreak of acute gastroenteritis in Anhui province was caused by genotype GII.4/Sydney 2012.
关键词
诺如病毒 /
G II.4基因型 /
Sydney 2012 /
分子特征
Key words
Norovirus /
G II.4 genotype /
Sydney 2012 /
Molecular characteristics
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基金
国家科技重大专项(2012ZX10004215)、公益性行业科研专项(20132004-1-2-2)
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