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ENGLISH ABSTRACT
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
王旌
朱剑锋
陆丽娜
作者及单位信息
·
DOI： 10.3760/cma.j.cn115989-20200430-00300
Classification of adenoviral conjunctivitis based on sequencing and phylogenetic analysis of hexon and fiber
Wang Jing
Zhu Jianfeng
Lu Lina
Authors Info & Affiliations
Wang Jing
Shanghai Eye Diseases Prevention & Treatment Center/Shanghai Eye Hospital, Shanghai General Hospital, National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Research Center of Precise Diagnosis and Treatment of Eye Diseases, Shanghai 200040, China
Zhu Jianfeng
Shanghai Eye Diseases Prevention & Treatment Center/Shanghai Eye Hospital, Shanghai General Hospital, National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Research Center of Precise Diagnosis and Treatment of Eye Diseases, Shanghai 200040, China
Lu Lina
Shanghai Eye Diseases Prevention & Treatment Center/Shanghai Eye Hospital, Shanghai General Hospital, National Clinical Research Center for Eye Diseases, Shanghai Key Laboratory of Ocular Fundus Diseases, Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai Engineering Research Center of Precise Diagnosis and Treatment of Eye Diseases, Shanghai 200040, China
·
DOI： 10.3760/cma.j.cn115989-20200430-00300
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摘要

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。

人类腺病毒;病毒性结膜炎;腺病毒六邻体蛋白;纤维蛋白;序列分析;系统发育
ABSTRACT

ObjectiveTo classify human adenovirus (HAdV) of adenoviral conjunctivitis by sequencing and phylogenetic analysis of hexon protein and fiber protein.

MethodsA 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.

ResultsAmong 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.

ConclusionsThe combined sequencing of hexon and fiber can provide abundant and effective biological information for the subtype and pathogenicity analysis of HAdV.

Adenoviruses, human;Conjunctivitis, viral;Hexon protein, adenovirus;Fiber protein;Sequence analysis;Phylogeny
Lu Lina, Email: mocdef.qabq910187anilul
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引用本文

王旌,朱剑锋,陆丽娜. 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[J]. 中华实验眼科杂志,2022,40(10)：902-907.

DOI:10.3760/cma.j.cn115989-20200430-00300

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腺病毒性结膜炎好发季节为春末至秋初，分为流行性角结膜炎(epidemic keratoconjunctivitis，EKC)、咽结膜热(pharyngeal conjunctival fever，PCF)和单纯滤泡性结膜炎，均可通过接触传播 [ 1 ]。EKC和PCF在密闭环境和高密度人群中极易传播和流行，甚至爆发。人类腺病毒(human adenovirus，HAdV)是EKC的主要病原体 [ 2 ]，其分型与腺病毒性结膜炎的分型、临床表现和预后密切相关。目前，基因鉴定技术已基本替代了传统血清学检测用于腺病毒检测，其中，六邻体蛋白是常见检测靶点，可对腺病毒进行初步分型，其次是纤维蛋白和五邻体蛋白，针对纤维蛋白的检测能够预测腺病毒的角膜致病性。与单基因测序相比，六邻体蛋白及纤维蛋白联合测序和系统发育分析能够更好地推断出HAdV分型，以及HAdV对角膜结膜上皮的黏附力和致病力 [ 3 ]。腺病毒多基因联合测序系统进一步的系统发育分析研究较少。本研究对六邻体蛋白和纤维蛋白进行联合测序及系统发育分析，以期提高腺病毒性结膜炎分型检测的敏感性，为准确预判流行特征、指导个性化用药提供新的方法。
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备注信息
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陆丽娜，Email： mocdef.qabq910187anilul
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王旌：实施研究、采集数据、统计分析、论文撰写和修改；朱剑锋、陆丽娜：分析数据、论文修改及定稿

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所有作者均声明不存在利益冲突
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上海市卫生和计划生育委员会青年课题项目 (20144Y0212)
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