腺病毒IgM抗体AlphaLISA快速检测方法

Establishment of a method for rapid detection of Ig M antibody against adenovirus by AlphaLISA

目的建立一种腺病毒IgM抗体的均相光激化学发光免疫快速检测方法(AlphaLISA)。方法通过筛选小鼠抗人IgM单克隆抗体标记受体微球、链霉亲和素偶联供体微球和生物素化腺病毒抗原的最适比例,构建腺病毒IgM抗体均相AlphaLISA快速检测体系;采用呼吸道感染样品对该检测体系进行评价,并与间接免疫荧光方法比较。结果与结论该法重复性较好,批内和批间变异系数均小于5%,与间接免疫荧光方法相比总符合率为87.21%,且不与其他常见呼吸道病原体的IgM抗体发生交叉反应。该法用于腺病毒IgM抗体的快速检测,可为早期确诊腺病毒感染提供可行方案。

Objective To establish a method for rapid detection of an IgM antibody against the adenovirus by homogeneous chemiluminescence immunoassay.Methods A homogeneous photoluminescence immunoassay system for adenovirus IgM antibodies was constructed by screening the optimal proportion of receptor beads labeled with mouse anti-human IgM monoclonal antibodies,streptavidin donor beads and biotinylated adenovirus antigens.The detection system was evaluated by respiratory tract infection samples and compared with the indirect immunofluorescence method.Results and Conclusion The repeatability of this method was good.The intra-assay and inter-assay coefficients of variations were less than 5%.Compared with the indirect immunofluorescence method,the total coincidence rate of this method was 87.21%,and there was no cross-reaction with IgM antibodies of other common respiratory pathogens.Therefore,this method could quickly detect IgM antibodies against adenovirus and provide an effective and feasible scheme for early diagnosis.