呼吸道感染人腺病毒的分离培养和型别鉴定  

Isolation,culture and identification of human adenovirus causing respiratory tract infection

作　　者：王瑞琨[1] 商蕾[1] 王迎晨[1] 金玉霞[1] 齐桂云[2] 黄冬娟[1] 曲章义[1] 

机构地区：[1]哈尔滨医科大学卫生微生物学教研室 [2]哈尔滨医科大学附属第二医院检验科,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2013年第5期385-387,F0004,共4页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目(81172725;30771909);教育部博士点专项基金(20070226007);黑龙江省自然科学基金重点项目(ZJY0701)

摘　　要：目的从临床呼吸道感染样品中分离人腺病毒毒株并进行型别鉴定。方法将经免疫荧光检测确认为人腺病毒感染的痰标本经处理后接种宫颈癌HeLa细胞进行培养,用空斑试验进行病毒分离和纯化。从纯化后的毒株中提取DNA作为模板,用PCR法扩增腺病毒六邻体高变区基因,对扩增产物进行核酸序列测定,测序结果运用基本局部比对搜索工具以确定其型别。结果从临床样品中分离到一株人腺病毒毒株,其六邻体基因高变区序列同人3型腺病毒六邻体基因的同源性为100%。结论从哈尔滨地区呼吸道感染患者痰标本中分离到人3型腺病毒一株。Objective To isolate the adenovirus from clinical sample taken from a patient with respiratory tract infection and to identify the type of the isolated adenovirus. Methods One adenovirus positive sample from hospitalized patients with respiratory tract infection was seeded onto HeLa cell. Adenovirus strain was isolated and purified from the virus plaque assay. The hypervarible region of hexon gene was amplified by the extracted viral DNA. via polymerase chain reaction （PCR）. The PCR product was sequenced and the sequence was subjected align- ment and basic local alignment search tool （BLAST） searching in GenBank. Results BLAST search analysis on Hexon gene showed that the adenovirus strain isolated was identical to human adenovirus type 3 strain USA/ak34_AdV3a2 with 100% homogeneity. Conclusion The ade- novirus isolated from a clinical sample is identified as human adenovirus type 3.

关 键 词：腺病毒 呼吸道感染 病毒培养 空斑试验 PCR 

分 类 号：R373.1[医药卫生—病原生物学]