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采用放射免疫测定法和酶免疫测定法从鼻咽标本中检测甲型流感病毒。

Detection of influenza A virus by radioimmunoassay and enzyme-immunoassay from nasopharyngeal specimens.

作者信息

Sarkkinen H K, Halonen P E, Salmi A A

出版信息

J Med Virol. 1981;7(3):213-20. doi: 10.1002/jmv.1890070305.

Abstract

Four-layer (indirect) radioimmunoassay (RIA) and enzyme-immunoassay (EIA) techniques were developed for the detection of influenza A and B virus in the sonicated nasopharyngeal specimens from patients hospitalized for acute respiratory infection. Polystyrene beads (RIA) or polystyrene microtiter plates (EIA) were used as the solid-phase, guinea pig antivirus immunoglobulins as the catching antibodies, rabbit antivirus immunoglobulins as the secondary antibodies, and 125I-labeled sheep antirabbit (RIA) or horseradish peroxidase conjugated swine antirabbit (EIA) immunoglobulins as the detector antibodies. A comparison of the developed RIAs and EIAs with the immunofluorescence (IF) method was made with 41 influenza A IF-positive and 150 influenza A IF-negative specimens. Each of the 41 influenza A IF-positive specimens was positive by the influenza A RIA and negative by the influenza B RIA. Out of 150 influenza A IF-negative specimens 3 specimens were found with weakly positive results in influenza A and B RIAs, but in each of these 3 specimens the binding proved nonspecific by the corresponding confirmatory tests. Using the EIa technique and the same immunoreagents as in RIA, identical results were obtained in each selected specimen tested. The developed RIAs and EIAs proved to be as specific and sensitive as the IF technique, and they should be practical in the diagnosis of respiratory infections directly from nasopharyngeal specimens.

摘要

开发了四层(间接)放射免疫分析(RIA)和酶免疫分析(EIA)技术,用于检测因急性呼吸道感染住院患者经超声处理的鼻咽标本中的甲型和乙型流感病毒。使用聚苯乙烯珠(RIA)或聚苯乙烯微量滴定板(EIA)作为固相,豚鼠抗流感病毒免疫球蛋白作为捕获抗体,兔抗流感病毒免疫球蛋白作为第二抗体,以及125I标记的羊抗兔(RIA)或辣根过氧化物酶偶联的猪抗兔(EIA)免疫球蛋白作为检测抗体。用41份甲型流感免疫荧光(IF)阳性标本和150份甲型流感IF阴性标本,将开发的RIA和EIA与免疫荧光(IF)方法进行了比较。41份甲型流感IF阳性标本中的每一份通过甲型流感RIA检测为阳性,通过乙型流感RIA检测为阴性。在150份甲型流感IF阴性标本中,有3份标本在甲型和乙型流感RIA中检测结果为弱阳性,但在这3份标本中的每一份中,相应的确证试验证明结合是非特异性的。使用EIA技术和与RIA相同的免疫试剂,在每个所选测试标本中获得了相同的结果。所开发的RIA和EIA证明与IF技术一样特异和灵敏,它们在直接从鼻咽标本诊断呼吸道感染方面应该是实用的。

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