摘要
建立了一种快速、灵敏检测牛乳中鼠伤寒沙门氏菌ATCC 14028(Salmonella typhimurium,S. typhimurium)的方法。利用鼠伤寒沙门氏菌多克隆抗体(Anti-S. typhimurium-pAb)修饰磁珠(Magnetic nanoparticles, MNPs)形成免疫磁珠(MNPs@pAb),鼠伤寒沙门氏菌单克隆抗体(Anti-S. typhimurium-mAb)标记多孔金@铂纳米酶(gold@platinum nanocatalysts,Au@PtNCs)形成具有催化性能的免疫探针Au@PtNCs@mAb,与鼠伤寒沙门氏菌最终反应形成MNPs@pAb-S. typhimurium-Au@PtNCs@mAb复合物后磁分离富集,在过氧化氢存在下可催化显色底物3,3’,5,5’-四甲基联苯胺(H2O2-TMB)生成蓝色物质,并在652 nm处产生较强的特征吸收峰。在最佳条件下,最低检测限为81 CFU/mL,线性范围为97~9.7×10^(5) CFU/mL,实际牛乳样本加标实验中加标回收率为96.75%~101.75%,变异系数(CV)为1.10%~5.13%。该方法建立的比色型生物传感器具有简单、快速、灵敏等特性,可用于检测乳品中的鼠伤寒沙门氏菌。
A rapid and sensitive method for the detection of Salmonella typhimurium(S.typhimurium)ATCC 14028 in milk was established.Using S.typhimurium polyclonal antibody(Anti-S.typhimurium pAb)to modify magnetic nanoparticles(MNPs)to form immune magnetic beads(MNPs@pAb),and using S.typhimurium monoclonal antibody(Anti-S.typhimurium-mAb)to label porous gold@platinum nanocatalysts(Au@PtNCs)to form immune probes Au@PtNCs@mAb with catalytic properties,which reacted with S.typhimurium to form MNPs@pAb-S.typhimurium-Au@PtNCs@mAb complexes and undergo magnetic separation and enrichment.In the presence of hydrogen peroxide,the chromogenic substrates 3,3',5,5'tetramethylbenzidine(H2O2-TMB)can be catalyzed to generate blue substances,and a strong characteristic absorption peak is generated at 652 nm.Under optimal conditions,the minimum detection limit is 81 CFU/mL,and the linear range is 97-9.7×10^(5) CFU/mL,the recovery rate of standard addition in the actual milk sample standard addition experiment is 96.75%-101.75%,and the coefficient of variation(CV)is 1.10%-5.13%.The colorimetric biosensor established by this method is simple,rapid and sensitive.It can be used to detect S.typhimurium in dairy products and has reference significance for food safety detection.
作者
苑懿
黄羽文
邢巾
张宇
YUAN Yi;HUANG Yuwen;XING Jin;ZHANG Yu(College of Food Science and Engineering Key Lab of Dairy Science,Ministry of Education,Northeast Agricultural University,Harbin 150030,China)
出处
《中国乳品工业》
CAS
北大核心
2023年第12期48-53,共6页
China Dairy Industry
关键词
多孔金@铂纳米酶
鼠伤寒沙门氏菌
比色
快速检测
porous gold@platinum nanocatalysts
S.typhimurium,colorimetry
rapid detection