基于六邻体蛋白及纤维蛋白序列测定的腺病毒性结膜炎分类分析

Classification of adenoviral conjunctivitis based on sequencing and phylogenetic analysis of hexon and fiber

目的基于六邻体蛋白和纤维蛋白的联合基因测序和系统发育分析进行腺病毒性结膜炎中人类腺病毒(HAdV)分型。方法在上海地区收集2015年1月至2017年8月病毒性结膜炎患者下结膜囊结膜拭子样本256份,提取DNA后通过PCR扩增六邻体蛋白及纤维蛋白全长,完成基因测序及种系发生学分析。结果89例(占34.76%)患者结膜拭子六邻体蛋白基因扩增阳性,包括HAdV-C1、HAdV-C2、HAdV-B3、HAdV-E4、HAdV-D8、HAdV-D19和HAdV-D37各1、7、20、6、23、17和15例,分别占1.12%、7.87%、22.47%、6.74%、25.84%、19.10%和16.85%,其六邻体蛋白种系发生学分析中均与相应参考序列原型聚合。纤维蛋白种系发生学分析中,15例HAdV-D19样本序列和1例HAdV-D37样本序列交叉聚合。结论针对六邻体蛋白和纤维蛋白基因的联合测序可以为HAdV分型和毒力分析提供丰富有效的生物学信息。

Objective To classify human adenovirus(HAdV)of adenoviral conjunctivitis by sequencing and phylogenetic analysis of hexon protein and fiber protein.Methods A total of 256 conjunctival swabs were collected from the inferior conjunctival sac of 256 patients with viral conjunctivitis in Shanghai from January 2015 to August 2017.After DNA extraction,the whole length of hexon and fiber was amplified by PCR to perform gene sequencing and phylogenetic analysis.The study protocol was approved by the Ethics Committee of Shanghai General Hospital(No.2020-202).Written informed consent was obtained from each subject.Results Among the samples,89(34.76%)were positive for hexon gene amplification,including 1(1.12%)of HAdV-C1,7(7.87%)of HAdV-C2,20(22.47%)of HAdV-B3,6(6.747%)of HAdV-E4,23(25.84%)of HAdV-D8,17(19.10%)of HAdV-D19,and 15(16.85%)of HAdV-D37.In phylogenetic analysis,sequenced hexon gene was clustered with the reference prototype correctly.In fiber phylogenetic analysis,15 of HAdV-D19 and 1 of HAdV-D37 were cross clustered.Conclusions The combined sequencing of hexon and fiber can provide abundant and effective biological information for the subtype and pathogenicity analysis of HAdV.