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Reagent kit for detecting antiuninuclear cell proliferation Listeria bacteria by colloidal gold Hly gene monoclonal antibodies

A technology of Listeria monocytogenes and monoclonal antibody, applied in the field of hygiene inspection and medical inspection, to achieve the effect of high titer, improved sensitivity and accuracy, and pure antibody

Inactive Publication Date: 2012-07-04
何鑫 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] 1 Specimen still needs 48h enrichment culture

Method used

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  • Reagent kit for detecting antiuninuclear cell proliferation Listeria bacteria by colloidal gold Hly gene monoclonal antibodies
  • Reagent kit for detecting antiuninuclear cell proliferation Listeria bacteria by colloidal gold Hly gene monoclonal antibodies
  • Reagent kit for detecting antiuninuclear cell proliferation Listeria bacteria by colloidal gold Hly gene monoclonal antibodies

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Experimental program
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Embodiment 1

[0058] Process conditions:

[0059] The first step is cloning and prokaryotic expression of the L. monocytogenes gene.

[0060] Main equipment:

[0061] Drugs and reagents: strains and plasmids Listeria monocytogenes strain (C54002 strain), pMD18-T, pGEX-6P vector plasmids are products of TAKARA company. Competent cells Escherichia coli JM109, BL21 recipient bacteria are products of Promega. Enzymes and related reagents Ex Taq polymerase, restriction endonucleases BamH Ⅰ, Xho Ⅰ and IPTG are products of Dalian Bao Biological Engineering Co., Ltd. T4 DNA ligase is a product of Promega. DNA Rapid Purification and Recovery Kit and Gel Rapid Purification and Recovery Kit are products of Dalian Bao Biological Engineering Co., Ltd.

[0062] experimental method:

[0063] 1. Genomic DNA preparation

[0064] Cultivate Listeria monocytogenes according to conventional methods, DNA rapid purification and recovery kit

[0065] Two. Containing the construction of L.monocytogenes Hly ...

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Abstract

The invention relates to a reagent kit for detecting antiuninuclear cell proliferation Listeria bacteria by colloidal gold Hly gene monoclonal antibodies and belongs to the technical field of hygiene inspection and medical inspection. The reagent kit is characterized by comprising the steps of: 1, primer design; 2, polymerase chain reaction (PCR) amplification; 3, agarose gel electrophoresis reaction conditions; 4, target deoxyribonucleic acid (DNA) segment cutting; 5, heat shock conversion method application; 6, reagent kit purification and plasmid preparation by a sodium dodecyl sulfate (SDS) cracking method; 7, protein induction expression; 8, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); 9, required strip cutting from the SDS-PAGE; and 10, colloidal gold preparation. The regent kit has the beneficial effect that the limitation aiming at the single-clostridium and single-antigen test in the prior art is avoided. The antibody is pure, the valence is high, and the sensitivity and the accurate degree are improved. The single added Listeria bacterium is detected. Compared with an enzyme-linked immuno sorbent assay (ELISA) method and a PCR method, the reagent kit has the advantage that the Listeria bacterium detection is faster and more convenient by the immune colloidal gold chromatography.

Description

technical field [0001] The invention belongs to the technical fields of hygienic inspection and medical inspection. Background technique [0002] Listeria is divided into 2 groups and 7 species. Group 1 includes Listeria monocytogenes L.monocytogenes (referred to as Listeria monocytogenes), Listeria ivanovii L.ivanovii, Listeria harmless L.innocua, Listeria westeri L.welshm ei and L.seeligeri; Group 2 is the less common L. grayi and L.m urrayi. Among them, L. monocytogenes is considered to be the cause of Listeria in animals and humans. The main pathogenic bacterium of terbiosis. Listeria monocytogenes (Usteria monocytogenes, Lm) is a kind of short Gram-positive bacillus, is a kind of pathogenic bacteria of zoonosis, can make people and animals sick Listeriosis. Studies have found that Listeria monocytogenes includes three types: pathogenic, weakly pathogenic and non-pathogenic. Listeria monocytogenes can cause diseases in humans and livestock, and cause death of pigs, ch...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531
Inventor 何鑫贾芙蓉黄胜起赵丽纯何成彦
Owner 何鑫
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