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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Clustal X 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。
Objective To understand the pathogen genotyping and molecular characteristics of an outbreak of acute gastroenteritis in a university in Anhui Province. Methods Real-time PCR was used to detect Norovirus nucleic acid in 6 samples of feces / anus swabs. The positive samples were amplified by RT-PCR to amplify part of the coat protein region (ORF2) Gene sequencing and type identification. Clustal X 2.0 and Mega 6.0 software were used to align the nucleotide sequences of the test sequences and construct phylogenetic relationship tree. Results 4 samples were detected positive for norovirus nucleic acid, the positive rate was 66.67% (4/6); gene sequence alignment and phylogenetic relationship showed that: Norovirus genotype GII.4; 2 strains of the sequence The nucleotide homology was 100%. The homology was 100% with Shanghai strain and Hong Kong strain in 2014-2015. The homology was 99.6% with 2012 Sydney strain (JX459908) The strain of kin in Hong Kong and the Mainland has recently become an independent cluster belonging to the GII.4 / Sydney 2012 branch. Conclusions The causative agent of the outbreak of acute gastroenteritis is Norovirus type GII.4 and belongs to the GII.4 Sydney 2012 variant.