鸡奇异变形杆菌外膜蛋白的抗血清制备及其荧光标记抗体检测方法的建立

Establishment of direct immunofluorescent assay for detection of Proteus mirabilis in chicken

为建立鸡奇异变形杆菌(P.mirabilis)直接荧光标记抗体检测方法,本研究提取P.mirabilis外膜蛋白(OMP),以OMP免疫健康新西兰家兔制备兔抗鸡P.mirabilis OMP高免血清,纯化的IgG,用FITC标记得到兔抗鸡P.mirabilis OMP-IgG的荧光标记抗体,建立直接荧光标记抗体检测方法。应用该方法检测临床样品184份,检测结果与细菌分离法和微量平板凝集比较,结果表明提取的OMP含有6种成分,分子量分别为67.0ku、63.1ku、57.5ku、53.7ku、43.0ku和31.6ku,经过优化的OMP-IgG荧光标记抗体最佳工作浓度为0.25mg/mL,最佳感作时间为30min。临床检测结果显示该方法具有简便、快速、敏感和特异性强等优点,为鸡P.mirabilis病的流行病学调查和临床快速诊断提供了技术保障。

A direct immunofluorescence assay for detection of Proteus mirabilis was developed using IgG antibody against the outer member protein （OMP）. The OMP-IgG was prepared from rabbit antiserum, purified and conjugated with FITC. The assay was optimized and could effectively detect P. mirabilis with 0.25 mg/mL OMP-IgG in 30 minutes. Test on 184 clinical samples showed that this assay was more sensitive and specific than the bacterial isolation method and microplate agglutination assay. It is simple to perform and could be used for rapid detection of P. mirabilis.