用免疫荧光方法检测鼻咽分泌物中多种呼吸道病毒抗原

Detection of multiple respiratory viral antigens in nasopharyngeal secretions with immunofluorescence assay

目的评价一种商品化的免疫荧光检测试剂（ＣｈｅｍｉｃｏｎＩｎｔｅｒｎａｔｉｏｎａｌＩｎｃ，Ｔｅｍｅｃｕｌａ，ＣＡ，ＵＳＡ）用于呼吸道病毒感染诊断的敏感性及特异性。方法间接免疫荧光方法，包括筛查及确定试验。筛查试剂中含有７种混合单克隆抗体，即甲型及乙型流感病毒、呼吸道合胞病毒（ＲＳＶ）、副流感病毒１、２、３型及腺病毒单克隆抗体。此外还有分别针对上述７种病毒的单克隆抗体用于确定试验。本研究用Ｃｈｅｍｉｃｏｎ试剂盒对９６例患儿的冻存鼻咽分泌物或支气管肺泡灌洗液细胞涂片标本进行回顾性检测，并与病毒分离结果进行比较。此外还用Ｃｈｅｍｉｃｏｎ试剂检测了１９９４年１１月～１９９５年２月北京儿童医院３００例下呼吸道感染患儿的鼻咽分泌物标本。结果与病毒分离结果相比，Ｃｈｅｍｉｃｏｎ确定试剂检测病毒抗原的敏感性为９５％、特异性为８６％。北京儿童医院的标本中有３８％（１０５／２８０）被确定为病毒感染，其中甲型流感病毒２０％、ＲＳＶ１４％、腺病毒３％、副流感病毒１、２、及３型７％。结论Ｃｈｅｍｉｃｏｎ呼吸道病毒诊断试剂能够同时检测多种呼吸道病毒抗原，对于确定呼吸道病毒感染是一种非常好的方法。但需要注意的是在筛查试验中有４％～８％的标?

Objective Antigen detection with immunofluorescence is an efficient method for diagnosis of respiratory tract infections, but such a method previously has not been applied for simple screening of multiple respiratory viruses Pools of monoclonal antibodies against various respiratory viruses are now available, and are proven to be potentially important tools for improvement of antigen detection in nasopharyngeal secretion specimen To evaluate the commercially available Chemicon immunofluorescence assay (IFA, respiratory viruses panel and identification kit), an indirect IFA containing a pool of monoclonal antibodies for screening for influenza virus types A and B, respiratory syncytial virus (RSV), parainfluenza virus types 1,2 and 3 and adenovirus, and the respective individual monoclonal ntibodies Methods Frozen nasopharyngeal secretions or bronchoalveolar lavage specimens from 96 patients with respiratory infections were retrospectively examined with the assay, and the results were compared with cell culture virus isolation Nasopharyngeal secretions from 300 children with lower respiratory tract infections collected at Beijing Childrens Hospital during the 19941995 winter season were also examined with the IFA Results The sensitivity of the Chemicon assay versus virus isolation was 95% and specificity 86% A viral etiology was identified in 105/280 (38%) Chinese children whose samples were evaluable (influenza A 20%, RSV 14%, adenovirus 3% and parainfluenza 1,2,3,7%) Conclusion The Chemicon IFA was shown to be an excellent tool for identification of viral respiratory tract infections although there was a problem with the the screening assay where around 4%8% of samples had nonspecific staining, necessitating verification test