Chemiluminescent Detect of E. coli O157:H7 Using Immunological Method Based on Magnetic Nanoparticles

文献类型： 外文期刊

第一作者： Li, Zhiyang

作者： Li, Zhiyang;He, Nongyue;Li, Song;Liu, Hongna;Li, Xiaolong;Li, Zhiyang;He, Lei;Shi, Zhiyang;Wang, Hua;Dai, Yabin;Wang, Zhifei

作者机构：

关键词： Magnetic Nanoparticles;Immunoassay;Alkaline Phosphatase;Chemiluminescence;Escherichia coli O157:H7

期刊名称：JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY （ 影响因子：1.134； 五年影响因子：0.999 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： The system of chemiluminescent magnetic enzyme-linked immunoassay was developed. E. coli O157:H7 was sandwiched between rabbits anti-E. coli O157:H7 polyclonal antibody-coated magnetic nanoparticles (immunomagnetic nanoparticles or IMNPs) and mouse anti-E. coli O157:H7 monoclonal antibody. Commercial alkaline phosphatase conjugated horse anti-mouse immunoglobulin (ALP-Ab) was used to bind with the monoclonal antibody, finally the chemiluminescent signals were detected by adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD) which was the substrate reagent of ALP. Different solvents of AMPPD were compared to get an optimal chemiluminescent signal. The effects of sodium borohydride and glycine on blocking the aldehyde groups of IMNPs were compared either, and the specificity and sensitivity of this system for detecting E. coli O157:1-17 were researched. The results indicated that Tris buffer was the best solvent of AMPPD, sodium borohydride was better than glycine in blocking IMNPs, and this method was of good specificity when using E. coli Top 10F' and Vibrio cholera as negative controls. The detection limit was 10~3 cells mL~(-1) when the antigen solution was 1 mL, and the procedure duration was about 3 h.

分类号： TB383

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