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用于快速灵敏检测流感病毒的乳胶比浊免疫分析法的开发

Development of latex turbidimetric immunoassay for rapid and sensitive detection of influenza virus.

作者信息

Nemoto Hiroshi, Komoriya Tomoe, Kohno Hideki

机构信息

Department of Applied Molecular Chemistry, College of Industrial Technology, Nihon University, 2-1 Izumi-cho 1-chome, Narashino-shi, Chiba 275-8575, Japan.

出版信息

Rinsho Biseibutshu Jinsoku Shindan Kenkyukai Shi. 2007;18(2):117-26.

Abstract

For the rapid and sensitive detection of influenza A and B viruses, a latex turbidimetric immunoassay (LTIA) was developed using latex reagents prepared by the sensitization of anti-influenza A or B monoclonal antibodies on latex particles. We measured the immunoreactivity of these latex reagents to influenza A and B viral antigens. The sensitivity and specificity of LTIA and reverse transcription-polymerase chain reaction (RT-PCR) for the detection of these viruses in clinical specimens (96 nasal swabs) were compared. The absorbance change in the latex agglutination reaction increased for each latex reagent with increasing concentration of the viral antigens. Reaction curves were obtained with each concentration of viral antigens for 5 min. The effective concentration ranges were 0-10 microg/ml for influenza A and 0-20 microg/ml for influenza B. The LTIA using clinical specimens revealed 8 positive and 73 negative results for influenza A and 15 positive and 52 negative results for influenza B. The sensitivities and specificities were 89% (8/9) and 84% (73/87), respectively, for influenza A and 100% (15/15) and 64% (52/81), respectively, for influenza B. The corresponding positive predictive values (PPV) were 36% (8/22) for influenza A and 34% (15/44) for influenza B. The negative predictive values (NPV) were approximately 99% (73/74) for influenza A and 100% (52/52) for influenza B. The LTIA is a rapid and sensitive method for detection of the influenza virus; It can be used for high throughput assay by automatic measurement and can potentially be used during influenza pandemics.

摘要

为了快速、灵敏地检测甲型和乙型流感病毒,我们利用抗甲型或乙型流感单克隆抗体致敏乳胶颗粒制备的乳胶试剂,开发了一种乳胶比浊免疫分析方法(LTIA)。我们测定了这些乳胶试剂对甲型和乙型流感病毒抗原的免疫反应性。比较了LTIA和逆转录聚合酶链反应(RT-PCR)在临床标本(96份鼻拭子)中检测这些病毒的敏感性和特异性。随着病毒抗原浓度的增加,每种乳胶试剂的乳胶凝集反应吸光度变化都有所增加。每种浓度的病毒抗原反应5分钟后获得反应曲线。甲型流感的有效浓度范围为0-10微克/毫升,乙型流感为0-20微克/毫升。使用临床标本的LTIA检测显示,甲型流感有8例阳性和73例阴性结果,乙型流感有15例阳性和52例阴性结果。甲型流感的敏感性和特异性分别为89%(8/9)和84%(73/87),乙型流感分别为100%(15/15)和64%(52/81)。甲型流感相应的阳性预测值(PPV)为36%(8/22),乙型流感为34%(15/44)。甲型流感的阴性预测值(NPV)约为99%(73/74),乙型流感为100%(52/52)。LTIA是一种检测流感病毒的快速、灵敏方法;它可通过自动测量用于高通量检测,并有可能在流感大流行期间使用。

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