副溶血弧菌高灵敏免疫层析检测法的建立

Establishment of a highly sensitive lateral-flow test method for the determination of Vibrio parahemolyticus

目的建立一种简便、准确、高灵敏的检测副溶血弧菌的免疫层析方法。方法采用免疫层析结合纳米颗粒信号放大技术,利用间接法进行示踪标记:试纸条由粘贴在底板上的样品垫、含有副溶血弧菌检测抗体的结合垫2、含有荧光颗粒标记羊抗鼠IgG的结合垫1、含有检测线(捕获抗体)和质控线(羊抗鼠IgG)的硝酸纤维素膜和吸水纸组成。结果结合增菌培养,该方法的检测灵敏性为10~3 CFU/g,与大肠杆菌、霍乱弧菌、河弧菌、创伤弧菌、溶藻弧菌、拟态弧菌、沙门氏菌、金黄色葡萄球菌等无交叉反应。结论本研究所建立的检测方法具有高灵敏、特异、简便、快速等优势,具有潜在的实际应用价值。

Objective To establish a simple, accurate and highly sensitive method for the detection of Vibrio parahemolytieus by lateral-flow test. Methods A signal amplification system was combined with lateral-flow test in order to improve the sensitivity, and the tracing mark was conducted by indirect method. The lateral-flow strip was constructed from a sample pad, 2 conjugated pads （one was coated with detection antibody, and the other was coated with fluorescence nanopartical-anti-mouse IgG conjugation）, a nitrocellulose membrane [containing detection line （capture antibody） and quality control line （sheep anti-rat IgG）] and an absorbent pad. Results The test strip had high sensitivity combined with pre-cultivation, and the sensitivity of this method was 10^3 CFU/g. This method had no cross reaction with E.coli, V.cholerae, V. flurialis, V.vulnificus, V.alginolyticus, V.mimicus, Salmonella spp. and Staphylococcus aureus. Conclusion This method is high sensitive, specific, simple, rapid, and has the potential application value in the rapid detection of Vibrio Parahemolyticus in food.