Study on Double-antibody Sandwich ELISA for Detecting E.coli O157 in Food

Abstract

Abstract: In this paper, anti-E.coli O157:H7 IgY was purified, and the titer of 10mg/ml purified IgY was 1:320. The anti- E.coli O157:H7 IgG was available by injection New Zealand rabbit with E.coli O157:H7 antigen, and the titer was 1:25600.Together with the IgY and IgG specific to E. coli O157, a double-antibody sandwich-ELISA was developed with IgG diluted 3200 fold as the capture antibody, and IgY as the detection antibody. Besides, the results of cross-experimentation of sandwich-ELISA described: ① IgG coating in 37℃ for 2h, ② reaction of IgG with antigen in 37℃ for 2h, and ③ concentration of IgY as 0.25mg/ml, and reaction of IgY with antigen in 37℃ for 1h. The detection limit of this method is 105CFU/ml, and has no cross reactions with other bacteria. With enrichment procedure in 41℃ and 200 r/min, E. coli O157:H7 recovers well from inoculated food samples. The results showed that the samples are detected after being cultured 12h at 0.1～1CFU/g(ml) and 8h at 1～ 10CFU/g(ml) respectively.

Key words: E. coli O157, double-antibody sandwich ELISA(enzyme linked immunosorbent assay), detect

GE Cui-Cui, ZHONG Qing-Ping, ZHANG Wang, 欧Yang-Xin . Study on Double-antibody Sandwich ELISA for Detecting E.coli O157 in Food[J]. FOOD SCIENCE, 2007, 28(1): 171-175.

[1]	LI Yining, HUANG Kunlun, YAO Zhiyi. Progress in the Construction and Application of Fluorescence Sensors for the Detection of Folic Acid [J]. FOOD SCIENCE, 2021, 42(9): 214-221.
[2]	NING Yawei, LIU Zhuo, YANG Zheng, MA Junmei, FAN Sufang, LI Qiang, ZHANG Yan. Simultaneous Detection of Three Major Milk Allergens in Food by UPLC-MS/MS [J]. FOOD SCIENCE, 2021, 42(8): 270-276.
[3]	WANG Yue, SHAO Lin, LI Qianxue, YI Le, QU Han, WANG Hongli, SHEN Minghao. Aptamer Combined with Quantum Dot for Simultaneous Detection of Staphylococcus aureus and Escherichia coli O157:H7 [J]. FOOD SCIENCE, 2021, 42(8): 305-312.
[4]	TIAN Jingsheng, LI Dongdong, ZHAO Lingyu, GAO Wenhui. Preparation and Application of Molecularly Imprinted Electrochemical Sensor for Rapid Detection of Quinoxaline Drug Residues [J]. FOOD SCIENCE, 2021, 42(6): 316-324.
[5]	CHEN Qiaoli, YANG Bing, HONG Qingyue, WEI Xunyu, FANG Chuchu, KAN Jianquan. Recent Progress in the Classification and Toxic Mechanism of Marine Biotoxins and Technologies for Their Detection [J]. FOOD SCIENCE, 2021, 42(5): 321-331.
[6]	LIU Na, CAO Qiang, XIAO Yushi, HUANG Rong, CHENG Lei, LIU Huan, HAN Gang, LI Jincheng, LI Menglong, WU Lidong. Recent Progress of Sensors Based on Dopamine and Its Derivatives in Food Safety Rapid Detection [J]. FOOD SCIENCE, 2021, 42(5): 332-339.
[7]	ZHAO Fei, CHEN Baoying, LI Kefeng, WANG Xu. Determination of Sialic Acid in Red Meat and Its Processed Products by High Performance Liquid Chromatography-Fluorescence Detection [J]. FOOD SCIENCE, 2021, 42(4): 313-318.
[8]	FU Ruijie, ZHOU Jing, TIAN Fengyu, JIAO Bining, HE Yue. Recent Progress in Nanomaterial-Based Optical Aptasensors for the Detection of Pesticide Residues [J]. FOOD SCIENCE, 2021, 42(3): 330-338.
[9]	WU Youxue, WU Meijiao, TIAN Yachen, WANG Zheng, GUO Liang, LIU Cheng, FANG Shuiqin, LIU Qing. Progress in the Development of Salmonella Biosensors [J]. FOOD SCIENCE, 2021, 42(3): 339-345.
[10]	WU Ying, FAN Congcong, SU Xiaona, SHEN Yudong, TAN Shu, ZHONG Cuili, XU Xiaoxuan, ZENG Daoping, YANG Jinyi. Simultaneous Detection of Four Veterinary Drugs Using Immunochip Assay Based on Smart Phone [J]. FOOD SCIENCE, 2021, 42(2): 278-282.
[11]	SHAN Shan, HUANG Zhaohong, HUANG Yanmei, LIU Daofeng, LIU Chengwei, HUANG Yunhong, LONG Zhong’er. Rapid Detection of Shiga Toxin-producing Escherichia coli by Immunochromatography Combined with Asymmetric Polymerase Chain Reaction [J]. FOOD SCIENCE, 2021, 42(2): 312-318.
[12]	HUANG Dongwei, GU Guizhang, HU Kena, GAO Xingjie, ZHANG Jinjie, YANG Wenge, XU Dalun. Detection of Guanine in Fish Flesh by Protein Adsorption onto Macroporous Resin Combined with Surface-Enhanced Raman Spectroscopy [J]. FOOD SCIENCE, 2021, 42(18): 298-305.
[13]	SHAN Shan, HUANG Yanmei, ZHAO Xuelong, CHEN Wenyao, LIU Chengwei, LONG Zhong’er, LIU Daofeng,. Detection of Escherichia coli O157:H7 by Cascade Signal Transduction System Combined with Immunochromatography [J]. FOOD SCIENCE, 2021, 42(18): 314-320.
[14]	YANG Jianfei, MA Qian, ZUO Yong, HUANG Xueqin, XU Jia, WANG Dingli, ZHOU Yaojin, YANG Keyi. A Spectrophotometric Reagent for the Rapid Detection of Nitrite and Its Application to Pickles [J]. FOOD SCIENCE, 2021, 42(18): 321-328.
[15]	CHEN Ruipeng, SUN Yunfeng, HUO Bingyang, QIN Yingkai, LI Shuang, LIANG Jun, ZHOU Huanying, GAO Zhixian. Progress in Multiple Detection Technologies for Mycotoxins [J]. FOOD SCIENCE, 2021, 42(17): 267-274.

Study on Double-antibody Sandwich ELISA for Detecting E.coli O157 in Food

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments