副溶血弧菌胶体金检测试纸条的改进

Improvement of colloidal gold based immunochromatographic strip test for detection of Vibrio parahaemolyticus

研制一种快速、灵敏、简便的胶体金免疫试纸条,用于改进副溶血弧菌的检测。通过胶体金与特异性针对于副溶血弧菌的单抗偶联,同时在NC膜上包被兔多抗、驴抗鼠的IgG分别作为检测线和质控线,制备免疫层系试纸条,测定试纸条的灵敏性与特异性。结果表明,当加入由胶体金标记的羊抗鼠二抗,能将显色强度提高5倍,明显有利于肉眼的观察。在纯培养物中试纸条的灵敏度为106CFU/mL,且与3株不同的副溶血弧菌均有阳性反应,与26株常见的菌均无交叉反应。同时在添加阪崎肠杆菌、志贺氏菌的鱼肉样本中,检测限为107CFU/mL。改良的副溶血弧菌胶体金免疫层系试纸条的建立,有助于加强食品中该菌的检测,提供了一种快速、灵敏、简便的方法。

To establish a rapid,specificity and sensitivity strip test for detection of Vibrio parahaemolyticus（VP）. Gold nanoparticles were coupled with monoclonal antibodies of VP,polyclonal antibodies against VP and Donkey anti-mouse IgG were separately coated on nitrocellulose membrane as test line and control line to detect the limit of detection and specifity. Results showed that color intensity increased by 5 times after adding the colloidal gold-labeled goat anti-mouse secondary antibody,was conducive to the naked eye observation. The detection sensitivity of the test was 10^6FU/mL in pure culture and 10^7CFU/mL in fish paste samples（in the presence of other kinds of bacteria species）. The strip had no cross-reaction with other bacterial strains, showing high specificity to VP. With a high specificity and sensitivity,the modified strip test is applicable in the detection of VP.