一种快速筛查金黄色葡萄球菌的荧光纳米免疫层析方法的建立  

Establishment of a fluorescent nanoparticle labeled immunochromatographic assay for the rapid detection of Staphylococcus aureus

作　　者：史咏梅[1] 何晖[2] 谭华[1] 冯子力[1] 伍碧梅[1] 朱海 涂承宁[1] 唐明慧[1] 杨泽[1] 汪海波[1] 

机构地区：[1]珠海国际旅行卫生保健中心,广东珠海519020 [2]广州市疾病预防控制中心 [3]深圳市易瑞生物技术有限公司

出　　处：《中国国境卫生检疫杂志》2015年第3期157-160,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：国家质检总局科技计划项目(2013IK248);广东省科技厅科研基金项目(2013B040402001)

摘　　要：目的建立一种快速筛查金黄色葡萄球菌的荧光纳米免疫层析检测方法。方法以荧光纳米颗粒作为示踪标记物,采用双抗体夹心法检测金黄色葡萄球菌A蛋白,制备了稀土离子标记的免疫层析试纸条,在紫外激发光源下判断反应结果。对该方法的特异性、敏感性和样本检测的适用性进行了分析。结果所制备的试纸条具有良好的特异性,与表皮葡萄球菌、腐生葡萄球菌、单核细胞增生李斯特菌、沙门菌、志贺菌、大肠杆菌、O1/O139群霍乱弧菌、副溶血性弧菌均无交叉反应。用该试纸条检测纯菌液和未经培养的粪便、呕吐物的灵敏度为7.2×104CFU/ml,检测经培养的模拟污染食物样本、粪便样本的检测限最低为7.2×102CFU/ml,反应在15 min内完成。结论本研究成功建立了金黄色葡萄球菌的荧光纳米免疫层析检测方法,能够快速检测食品、奶制品、呕吐物、粪便中的金黄色葡萄球菌,具有良好的特异性和灵敏性,便于开展现场快速检测。Objective To develop a fluorescent nanoparticle labeled immunochromatographic strip for the rapid detection of Staphylococcus aureus. Methods Fluorescent nanoparticles were conjugated with monoclonal antibodies against Staphylococcus aureus and then employed in the development of the immunochromatographic strip in a sandwich format. Reaction results were detected under UV. The specificity and sensitivity, as well as the clinical application of the strip were evaluated. Results The strip showed no cross-reaction with Staphylococcus epidermidis,Saprophytic Staphylococcus, Listeria monocytogenes, Salmonella enteritidis, Shigella flexneri, Escherichia coli, Vibrio cholerae O1/O109, Vibrio parahaemolyticus, indicating its high specificity. The sensitivity of the strip was 7.2 ×10^4CFU/m L when cultured bacterial liquid, uncultured vomit or stool samples were tested. The limit of detection could reach as low as 7.2×10^2CFU/m L when artificial contaminant milk was detected. The detection can be finished in 15 min and was applicable to food, milk, vomit and stool samples. Conclusion The fluorescent nanoparticle labeled immunochromatographic strip was established successfully. This method showed high sensitivity and specificity for the rapid detection of Staphylococcus aureus in food, milk, vomit and stool, which was valuable for the detection in locale.

关 键 词：荧光纳米颗粒 免疫层析试纸条 金黄色葡萄球菌A蛋白 

分 类 号：R117[医药卫生—公共卫生与预防医学]