腺病毒六邻体蛋白的基因测序在流行性角膜结膜炎病原检测中的应用  

Application of hexon gene sequencing of adenovirus in pathogen detection of epidemic keratoconjunctivitis

作　　者：王旌[1] 赵蓉[1] 童晓维[1] 朱剑锋[1] 

机构地区：[1]上海市眼病防治中心,200040

出　　处：《中华实验眼科杂志》2015年第5期430-435,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：上海市卫生局青年课题项目（2010Y135）

摘　　要：背景 流行性角膜结膜炎是常见的眼部传染性疾病,腺病毒为常见病原体之一.六邻体蛋白是构建病毒衣壳的重要蛋白,也是病毒与抗体结合的主要靶点.应用针对六邻体蛋白编码区的测序方法可对腺病毒进行快速分型,对于流行性角膜结膜炎的早期诊断、病情监测和预后判断具有重要意义. 目的 通过针对腺病毒六邻体蛋白编码区的测序,完成上海地区腺病毒与流行性角膜结膜炎相关的分子流行病学调查.方法 在受检者知情同意的情况下,收集2010年1月至2012年12月在上海市眼病防治中心和上海市急性出血性结膜炎防控办公室下属红眼病监测站点就诊的214例可疑流行性角膜结膜炎患者结膜囊的结膜拭子样本214份.对结膜囊标本提取DNA后,通过PCR法扩增病原体六邻体蛋白编码区内140 bp的保守序列,对于腺病毒阳性的标本,通过对六邻体蛋白保守区域内956 bp的序列进一步测序以确定其病毒分型. 结果 在214例患者中,109例在通用PCR中显示腺病毒阳性,占50.93％.其中通过基因测序确定AdV1者4例,AdV2者33例,AdV3者15例,AdV4者12例,AdV8者19例,AdV19者15例,AdV37者8例.属于D组腺病毒的AdV8、AdV19和AdV37检出阳性患者更容易出现角膜炎症反应、伪膜性结膜炎和耳前淋巴结肿大等,而B组腺病毒检出阳性者易伴发上呼吸道感染,但角膜受累者较少.结论 针对六邻体蛋白编码区内保守区域的测序是腺病毒分型的重要方法之一,可快速确定流行性角膜结膜炎是否系腺病毒感染引起,并确定腺病毒分型.Background Epidemic keratoconjunctivitis is a common eye disease,and adenovirus is one of the common pathogens.The hexon protein,one main capsid protein of the virus,is an important target of antibody binding.Thus,sequencing the coding region of the hexon protein is an important way for adenovirus fast typing.Objective This study was to complete a molecular epidemiology survey of epidemic keratoconjunctivitis and investigate its association with adenovirus in Shanghai area by sequencing the coding region of hexon protein.Methods Two hundred and fourteen sacconjunctival swab specimens were collected from 214 patients with suspicious epidemic keratoconjunctivitis who visited Shanghai Eye Disease Prevention and Treatment Center and the clinical sites supervised by the Shanghai Prevention and Monitoring Office of Acute Hemerragic Conjunctivitis under the informed consent from January 2010 to December 2012.DNA was extracted from the specimens and then the 140 bp conserved sequence in hexon protein coding region was amplified by PCR initially to determine an adenovirus pathogen.Furtherly,956 bp conserved sequence of the hexon codind district was sequencied to clarify the serotype of adenovirus in the adenovirus-positive specimens.Results 50.93% patients （109/214） were detected to be adenovirus-positive by generic PCR,in which AdV1 ＋ was in 4 patiens,AdV2＋ was in 33 patients,AdV3＋ was in 15 patients,AdV4＋ was in 12 patients,AdV8＋ was in 19 patiens,AdV19＋ was in 15 patients,AdV37＋ was in 8 patients.The subgenus D adenoviruses,including AdV8＋,AdV19＋ and AdV37＋ often resulted in corneal inflammation,pseudomembranous conjunctivitis and preauricular lymph nodes;while subgenus B adenovirus induced much frequent tract infection and less corneal response.Conclusions PCR-sequence of conserved region of hexon protein coding district is applicable for the detection and serotyping of adenovirus in epidemic keratoconjunctivitis.

关 键 词：人腺病毒感染 病毒性角膜结膜炎/诊断 碱基序列 保守序列 PCR/方法 六邻体蛋白 

分 类 号：R772.21[医药卫生—眼科]