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急性呼吸道感染的快速病毒诊断:酶联免疫吸附测定法与免疫荧光技术检测鼻咽分泌物中病毒抗原的比较

Rapid viral diagnosis of acute respiratory infections: comparison of enzyme-linked immunosorbent assay and the immunofluorescence technique for detection of viral antigens in nasopharyngeal secretions.

作者信息

Grandien M, Pettersson C A, Gardner P S, Linde A, Stanton A

出版信息

J Clin Microbiol. 1985 Nov;22(5):757-60. doi: 10.1128/jcm.22.5.757-760.1985.

Abstract

Nasopharyngeal secretions from adults and children were obtained in Stockholm, Sweden, for routine diagnosis of influenza A virus, influenza B virus, respiratory syncytial (RS) virus, parainfluenza type 3 virus, and adenovirus infections by demonstration of viral antigens directly in the specimens. The cells in nasopharyngeal secretions were pelleted by centrifugation for preparation of cell deposits for diagnosis by the immunofluorescence technique (IF) in London, England, and in Stockholm, whereas the supernatants were used to diagnose infection by the enzyme-linked immunosorbent assay (ELISA) in Stockholm. Titrations of the various purified viruses showed that ELISA could detect viral antigens in amounts corresponding to 1 to 10 ng of virus protein per test well. In a series of 73 specimens tested for influenza A, RS, and parainfluenza type 3 viruses by IF in London and by ELISA in Stockholm, 15 of 18 RS, 14 of 15 influenza A, and 2 of 2 parainfluenza type 3 viral infections were diagnosed by ELISA as compared with IF, giving sensitivities for RS and influenza A viral diagnosis of 83 and 93%, respectively, and a specificity of 100%. In another series of specimens from 35 patients tested for influenza B virus and adenovirus, five influenza B virus and four adenovirus infections were diagnosed by both methods; one additional influenza B infection was detected only by IF and another only by ELISA. Comparisons of diagnostic results between the two methods performed in Stockholm gave nonagreement of results for 37 of 1,593 tests (2.5%) for the five viruses. The conclusion reached was that the described ELISA, although a satisfactory test, had somewhat less sensitivity than did IF for the detection of respiratory viral infections. This could possibly be explained by unnecessary dilutions of specimens at the time of collection; transportation, processing, and storage of specimens were less complicated than for IF.

摘要

在瑞典斯德哥尔摩采集成人和儿童的鼻咽分泌物,用于通过直接在标本中检测病毒抗原来常规诊断甲型流感病毒、乙型流感病毒、呼吸道合胞(RS)病毒、3型副流感病毒和腺病毒感染。鼻咽分泌物中的细胞通过离心沉淀,以便在英国伦敦和斯德哥尔摩用免疫荧光技术(IF)制备用于诊断的细胞沉淀物,而在斯德哥尔摩,上清液用于通过酶联免疫吸附测定(ELISA)诊断感染。各种纯化病毒的滴定表明,ELISA能够检测到每个测试孔中相当于1至10 ng病毒蛋白量的病毒抗原。在伦敦通过IF和在斯德哥尔摩通过ELISA对73份检测甲型流感、RS和3型副流感病毒的标本进行的一系列检测中,与IF相比,ELISA诊断出18份RS感染中的15份、15份甲型流感感染中的14份以及2份3型副流感病毒感染中的2份,RS和甲型流感病毒诊断的敏感性分别为83%和93%,特异性为100%。在另一组来自35名患者检测乙型流感病毒和腺病毒的标本中,两种方法均诊断出5例乙型流感病毒感染和4例腺病毒感染;仅通过IF检测到另外1例乙型流感感染,仅通过ELISA检测到另外1例。在斯德哥尔摩对两种方法的诊断结果进行比较,对于这五种病毒,1593次检测中有37次(2.5%)结果不一致。得出的结论是,所述的ELISA虽然是一种令人满意的检测方法,但在检测呼吸道病毒感染方面的敏感性略低于IF。这可能是由于标本采集时不必要的稀释所致;标本的运输、处理和储存比IF简单。

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