摘要
为提高食品中副溶血弧菌、创伤弧菌和拟态弧菌毒素初筛的速度与效率,建立了双抗体夹心ELISA检测方法。以融合蛋白TDH-VVC-VMHA包涵体为抗原免疫豚鼠,采用豚鼠抗融合蛋白免疫血清为包被抗体,辣根过氧化物酶(HRP)标记羊抗豚鼠IgG为标记抗体,应用棋盘滴定法确定ELISA的最适条件。结果表明,所建立的方法与副溶血弧菌、创伤弧菌、拟态弧菌的培养物上清有较明显的阳性反应,与绿脓杆菌、金黄色葡菌球菌、李氏杆菌、甲型副伤寒沙门氏菌、猪霍乱沙门氏菌等5种非弧菌属食物中毒菌培养物上清未见交叉反应,与溶藻弧菌、河流弧菌和麦氏弧菌等3种弧菌培养物上清也未见交叉反应。这说明,该方法可以用于食品中副溶血弧菌、创伤弧菌和拟态弧菌毒素的快速同步免疫学检测。
In order to improve the detection speed and efficiency,a double antibody sandwich ELISA was developed to detect Vibrio parahaemolyticus,Vibrio vulnificus or Vibrio minicus in food simultaneously.The cytorrhyctes of the fusion protein were purified by the electroeluting to immunize cavia cobaya,anti-serum were uesed as capture antibody,goat anti-guinea pig IgG/HRP were uesed as labelled-antibody,optimizing condition was determined by use of the chessboard titration method.The results showed that,the double antibody sandwich ELISA having obvious positive reaction with the culture supernatant of Vibrio parahaemolyticus,Vibrio vulnificus,Vibrio mimicus,but no positive reaction with 8 other culture supernatant of contrast strains including Pseudomonas aeruginosa,Staphylococcus aureus,Listeria,etc.It indicated that this method could be used to detect Vibrio parahaemolyticus,Vibrio vulnificus and V.mimicus toxin in food simultaneously.
出处
《中国农学通报》
CSCD
北大核心
2011年第32期34-37,共4页
Chinese Agricultural Science Bulletin
基金
辽宁省教育厅科研项目"三种食源中毒性弧菌融合毒素的基因表达及ELISA检测方法的建立"(L2010262)
