Double-antibody sandwich method for detecting salmonella typhimurium in food based on monoclonal antibodies

A monoclonal antibody, double antibody sandwich technology, applied in the field of immunoassay, can solve the problems of weak LPS immunogenicity, difficulty in monoclonal antibody, and difficulty in causing immune response, and achieves good stability, high uniformity, and simple pretreatment process. Effect

Active Publication Date: 2014-02-05
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The successful preparation of antibodies is inseparable from the immunogen used, and it is difficult to prepare monoclonal antibodies against Salmonella simply through bacterial immunization
As the main antigen of Salmonella, LPS is a potential detection antigen, but the immunogenicity of LPS is weak, and it is difficult to induce sufficient immune response in mice

Method used

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  • Double-antibody sandwich method for detecting salmonella typhimurium in food based on monoclonal antibodies

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Embodiment Construction

[0037] specific implementation plan

[0038] The present invention is further illustrated by the following examples.

[0039]1. Instrument:

[0040] TGL-40B desktop low-speed centrifuge, Shanghai Anting Scientific Instrument Factory

[0041] KFLOW pure water machine, KFLOW company

[0042] ZD-9556 horizontal shaker, Taicang Science and Education Equipment Factory

[0043] 96-well 8×12 detachable ELISA plate, Xiamen Yijiamei Laboratory Equipment Co., Ltd.

[0044] MuLtiska Mks Microplate Reader, Thermo Labsystems

[0045] Adjustable pipette, Thermo Labsystems

[0046] Vortex mixer, Shanghai Huxi Instrument Analysis Factory

[0047] 2. Reagents:

[0048] Tetramethylbenzidine (TMB), Shanghai Jingchun Industrial Co., Ltd.

[0049] Other reagents are analytical reagents

[0050] 3. Steps

[0051] 1. Preparation of monoclonal antibodies

[0052] (1) Experimental animals: 5 7-week-old BALB / c mice were selected for immunization.

[0053] (2) Antigen configuration: Dilute ...

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Abstract

The invention discloses a double-antibody sandwich method for detecting salmonella typhimurium in food based on monoclonal antibodies, belonging to the technical field of immunoassay. Salmonella typhimurium ATCC13311 and smooth salmonella typhimurium LPS are adopted for mixed immunity of a 7-week BALB / c mouse, 10 LPS monoclonal antibodies are obtained by immunity, fusion and screening, horse radish peroxidases (HRP) are labeled respectively, and the salmonella typhimurium is paired two by two. A sandwich enzyme-linked immuno sorbent assay (ELISA) method is established by taking 6E2 CGMCC No.7206 monoclonal antibodies as coated antibodies and enzyme-labeled antibodies and by taking the salmonella typhimurium as standards, and the LOD is 500cfu / mL. The sandwich method, established by using the monoclonal antibodies which are highly uniform in physicochemical property and high in specificity and can be prepared on a large scale, is high in sensitivity and low in cost; the salmonella typhimurium is not in cross reaction with salmonella enteritidis, salmonella arizonae, E.coli, E.coliO157:H7, enterobacter sakazakii, staphylococcus aureus and listeria monocytogenes; a quick and efficient analysis way is provided for detection of the salmonella typhimurium in the food.

Description

technical field [0001] The invention relates to a double-antibody sandwich method for detecting Salmonella typhimurium in food based on a monoclonal antibody, which belongs to the technical field of immune analysis. Background technique [0002] Salmonella is a global foodborne pathogen. Biologically, Salmonella is a kind of Gram-negative bacteria with blunt ends, no spores, and generally no capsule. The main antigens are O antigen, H antigen, and Vi antigen. Animal foods such as poultry, eggs, and dairy products are easily contaminated with Salmonella. After the human body ingests food containing bacteria, it will cause acute gastroenteritis, typhoid fever, children with low immunity and even sepsis and other symptoms. [0003] There are more than 2,000 serotypes of Salmonella, and the common serotypes in clinical practice are mainly Salmonella enteritidis, Salmonella typhimurium, and Salmonella paratyphi A. The application of well-regulated production operations and man...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
CPCG01N33/531G01N33/56916G01N33/577
Inventor 胥传来王文彬匡华徐丽广马伟刘丽强
Owner JIANGNAN UNIV
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