牛奶模拟样本中金黄色葡萄球菌肠毒素C含量的两种发光检测方法比较

Comparison of two luminescence detection methods for staphylococcal enterotoxin C content in simulated milk samples

目的 比较均相光激化学发光检测法(AlphaLISA)和磁微粒化学发光检测法(MP-CLIA)在检测牛奶模拟样本中金黄色葡萄球菌肠毒素C(SEC)含量的敏感性和精确性。方法 以山羊抗SEC多克隆抗体偶联受体微球、生物素标记的SEC单克隆抗体和链霉亲和素偶联的供体微球,构建AlphaLISA检测体系;以山羊抗SEC多克隆抗体偶联碱性磷酸酶、生物素标记的SEC单克隆抗体和链霉亲和素偶联磁珠,构建MP-CLIA检测体系。结果 AlphaLISA检测牛奶模拟样本中SEC含量的灵敏度为4.04 ng/L,变异系数(CV)为1.98%~9.82%;MP-CLIA的灵敏度为108.19 ng/L,CV为4.63%~20.40%。结论 与MP-CLIA相比,AlphaLISA检测牛奶模拟样本中SEC含量的敏感性和精确性更高。

Objective To compare the sensitivity and accuracy of amplified luminescent proximity homogeneous assay linked immunosorbent assay(AlphaLISA)and magnetic particles-based chemiluminescence immunoassay(MP-CLIA)for detection of staphylococcal enterotoxin C(SEC)in the simulated milk samples.Methods The AlphaLISA was constructed using goat anti-SEc polyclonal antibody-coupled receptor microspheres,biotin-labeled SEC monoclonal antibody and streptavidin-coupled donor microspheres.The MP-CLIA was constructed using goat anti-SEC polyclonal antibody conjugated alkaline phosphatase,biotin-labeled anti-SEC monoclonal antibody and streptavidin conjugated magnetic beads.Results The sensitivity of AlphaLISA to detect SEC content in simulated milk samples was 4.04 ng/L,and the coefficient of variation(CV)was 1.98%~9.82%.The sensitivity of MP-CLIA was 108.19 ng/L and CV was 4.63%~20.40%.Conclusion Compared with MP-CLIA,AlphaLISA is more sensitive and accurate to detecting SEC.