Rapid Detection of Six Salmonella Serotypes in Chilled Fresh Meat by Double Antibody Sandwich ELISA

doi:10.7506/spkx1002-6630-201616034

FOOD SCIENCE

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Rapid Detection of Six Salmonella Serotypes in Chilled Fresh Meat by Double Antibody Sandwich ELISA

ZHANG Shuai, QI Yingying, ZHANG Hongxing*, WANG Yiwen, XIE Yuanhong, LIU Hui

Beijing Laboratory of Food Quality and Safety, Beijing Key Laboratory of Agricultural Product Detection and Control for Spoilage
Organisms and Pesticide, Beijing Engineering Technology Research Center of Food Safety Immune Rapid Detection,
Beijing Engineering Laboratory of Key Technology Development of Microecologics, College of Food Science and Engineering,
Beijing University of Agriculture, Beijing 102206, China

Online:2016-08-25 Published:2016-08-30
Contact: ZHANG Hongxing

Abstract

Abstract:

This study aimed to prepare antibodies specific for Salmonella and to use them establish an enzyme linked
immunosorbent assay (ELISA) system for the rapid detection of six Salmonella serotypes in chilled fresh meat. In this
study, BALB/c mice and rabbits were immunized with mixed antigens from six pathogenic strains of Salmonella for the
production of monoclonal antibodies using hybridoma technology for cell fusion. The results showed that a hybridoma
cell line (6E7 CGMCC 10313) which could stably secret monoclonal antibody (McAb, M23) specific for Salmonella was
successfully screened and the polyclonal antibody (P23) was also obtained at the same time. The titer of the prepared highly
pure monoclonal and polyclonal antibodies was 1:1.28 × 106 and 1:8.0 × 105, respectively. P23 was adsorbed on 96-well
microtiter plates as coating antibody and M23 was labeled with horseradish peroxidase for the establishment of sandwich
ELISA. The limit of detection (LOD) for Salmonella in artificially contaminated meat samples was 800 CFU/g. There was
no cross-reactivity with Shigella, Enterobacter sakazakii, E. coli O157:H7, Staphylococcus aureus, Listeria monocytogenes
and other Salmonella.

Key words: Salmonella, hybridoma technology, monoclonal antibody, sandwich ELISA

CLC Number:

TS251.7

ZHANG Shuai, QI Yingying, ZHANG Hongxing, WANG Yiwen, XIE Yuanhong, LIU Hui. Rapid Detection of Six Salmonella Serotypes in Chilled Fresh Meat by Double Antibody Sandwich ELISA[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201616034.

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Rapid Detection of Six Salmonella Serotypes in Chilled Fresh Meat by Double Antibody Sandwich ELISA

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