快速灵敏检测流感病毒的乳胶比浊免疫分析法的建立。

Hiroshi Nemoto, Tomoe Komoriya, Hideki Kohno
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引用次数: 0

摘要

为了快速、灵敏地检测甲型流感病毒和乙型流感病毒,建立了一种乳胶浊度免疫分析法(LTIA),该方法采用抗甲型流感病毒或乙型流感病毒单克隆抗体在乳胶颗粒上致敏制备乳胶试剂。我们测量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LTIA和RT-PCR检测临床标本(96份鼻拭子)的敏感性和特异性。随着病毒抗原浓度的增加,胶乳凝集反应的吸光度变化增大。不同浓度的病毒抗原反应曲线分别为0 ~ 10 μ g/ml和0 ~ 20 μ g/ml。临床标本LTIA检测结果显示,甲型流感阳性8例,阴性73例,乙型流感阳性15例,阴性52例,敏感性和特异性分别为89%(8/9)和84%(73/87),甲型流感和100%(15/15)和64%(52/81)。甲型流感和乙型流感的阳性预测值分别为36%(8/22)和34%(15/44),甲型流感和乙型流感的阴性预测值分别为99%(73/74)和100%(52/52)。LTIA是一种快速、灵敏的流感病毒检测方法;它可以通过自动测量用于高通量分析,并可能在流感大流行期间使用。
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Development of latex turbidimetric immunoassay for rapid and sensitive detection of influenza virus.

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.

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