胶体金免疫层析法检测猪链球菌2型的研究

Development of Colloid Gold Immunochromatography for Rapid Detection of Streptococcus suis Serotype 2

目的:制备胶体金免疫层析试纸检测猪链球菌2型。方法:用柠檬酸盐还原法制备胶体金颗粒,标记猪链球菌2型多克隆抗体,通过免疫层析作用对猪链球菌2型进行检测,并对试纸条的敏感性、特异性、稳定性进行评价。结果:每毫升胶体金最佳抗体标记量为22μg/mL,最佳包被抗体浓度为2mg/mL,最佳BSA封闭浓度为1.5%,建立的胶体金免疫层析试纸条检出猪链球菌2型的下限为106CFU/mL,从检测到结果判断时间为5～15min,与其他常见致病菌及链球菌属中15个群无交叉反应。结论:获得了检测猪链球菌2型的胶体金免疫层析试纸,该法操作简便,灵敏度高,特异性强,可用于猪链球菌的快速初筛和检测。

Objective： To prepare colloidal gold dipstick for rapid detection of Streptococcus suis serotype 2. Methods： Colloidal gold was prepared by citrate-natrium deoxidize method and coupled with the antibody against Streptococcus suis serotype 2, and Streptococcus suis serotype 2 was detected by immunochromatography assay in this study. Besides its sensitivity, specificity and stability were studied. Results： Gold immunochromatography assay was developed for detection of Streptococcus suis serotype 2, the best compatible protein concentration was measured as 22 μg/mL, the best coating protein concentration was measured as 2 mg/mL and the best blocking concentration was 1.5% of BSA. The detection sensitivity was as low as 106 CFU/ml and the strip has no cross-action with other bacteria detected. It took only 5-15 min from detecting to judging the result. Conclusions： The colloidal gold dipstick was prepared successfully, which was feasible, convenient, quick, and effective for detection of Streptococcus suis serotype 2.