近年北京儿科急性呼吸道感染患儿中腺病毒的型别监测  

Identification and typing of adenoviruses from pediatric patients with acute respiratory infections in Beijing from 2003 to 2008

作　　者：邓洁[1] 钱渊[1] 赵林清[1] 朱汝南[1] 王芳[1] 孙宇[1] 廖斌[2] 黄荣妍[2] 袁艺[2] 曲东[2] 任晓旭[2] 

机构地区：[1]首都儿科研究所病毒研究室北京市感染与免疫中心实验室,北京100020 [2]首都儿科研究所附属儿童医院

出　　处：《中华儿科杂志》2010年第10期739-743,共5页Chinese Journal of Pediatrics

基　　金：国家自然科学基金（30872153）

摘　　要：目的 了解近年北京地区儿科急性呼吸道感染患儿中人腺病毒（ADV）感染状况及ADV的型别.方法 2003年至2008年,连续6年通过病毒分离和（或）间接免疫荧光法,对收集的首都儿科研究所就诊的急性呼吸道感染患儿的17 941份标本进行ADV检测,对其中285份毒株或阳性标本提取DNA,通过3、7、11和21型4对分型引物的多重聚合酶链反应,确定3、7、11和21型ADV,对少数扩增阴性的标本或毒株,再使用2对可扩增所有ADV的通用引物对其DNA进行扩增,将PCR产物直接测序,结果与GenBank上的序列比较,确定其型别.结果 在17 941份呼吸道标本中,经病毒分离和（或）间接免疫荧光法确定为ADV阳性的标本304份,总阳性率为1.69%.对其中285份毒株或阳性标本的PCR分型结果：272例用3、7、11、21型分型引物扩增确定了型别：3型ADV 174例,占61.1%（174/285）,7型ADV 92例,占32.3%,11型ADV 6例,占2.1%;有13例经分型引物扩增为阴性,经过通用引物扩增后的PCR产物测序确定为2型ADV 9例,占3.2%（9/285）;6型ADV2例,占0.7%,1型和5型ADV各1例,各占0.4%.结论 本组ADV阳性检出率为1.69%.近年来北京地区ADV感染以3、7型为主,其次为2型和11型,1、5和6型较为少见,尚未发现其他型别.Objective Adenovirus （ADV） is one of the most common causes of acute respiratory infections in infants and children. The objective of this study was to investigate the prevalence of adenovirus infection among pediatric patients with acute respiratory infections in Beijing and the types of the adenoviruses circulating in Beijing on the molecular bases. Method Clinical specimens including throat swabs from outpatients and nasopharyngeal aspirates from hospitalized patients were collected from patients with acute respiratory infections in a consecutive period of 6 years from Jan 2003 to Dec 2008. Adenoviruses were identified from the collected clinical specimens by tissue culture and/or immunofluorescence assay and typed by nested-PCR based on the sequence of the encoding gene of hexon. Primers were designed for PCR amplification using hexon gene of adenovirus as target. One primer pair was designed as universal primers for amplifying a 1278 bp gene fragment located at the hexon gene of adenovirus types 3, 7, 11 and 21. Four primer pairs with the sequences located within the region of this 1278 bp fragment were designed specifically for amplifying adenoviruses types 3, 7, 11 or 21, respectively, which were used for a multiplex nest-PCR in a single tube. The products from this multiplex nest-PCR were 502 bp （for type 3）, 311 bp （for type 7）,880 bp （ for type 11 ） and 237 bp （ for type 21 ）, respectively, and the type of the adenovirus tested can be determined after agarose electrophoresis analysis of the PCR products. For those strains which could not be typed by the multiplex nest-PCR, the gene fragment was amplified by a universal primer pair for all adenovirus types from group A to F and the PCR products were sequenced directly. Result Out of 17 941clinical specimens collected, including 4378 throat swabs from outpatients and 13 563 nasopharyngeal aspirates from hospitalized patients, 304 were adenovirus positive by tissue culture and/or immunofluorescence assay, the overall positive rate was 1

关 键 词：腺病毒 人 腺病毒感染 人 呼吸道感染 儿童 

分 类 号：R686[医药卫生—骨科学]