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Colloidal gold labeling lap gene monoclonal antibody measuring Listeria monocytogenes kit

A technology of Listeria monocytogenes and monoclonal antibodies, which is applied in the field of hygienic inspection and medical inspection, and can solve the problem of incompleteness of the entire genus of Listeria

Inactive Publication Date: 2009-12-30
何成彦 +3
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The antibody used by Luo Lixin and others is Listeria monocytogenes antibody (O chain antigen I / II). The experiment was only conducted on a single serotype of Listeria. and flagellar (H) antigens are divided into 13 serotypes, so the experiment is not complete for Listeria

Method used

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  • Colloidal gold labeling lap gene monoclonal antibody measuring Listeria monocytogenes kit
  • Colloidal gold labeling lap gene monoclonal antibody measuring Listeria monocytogenes kit
  • Colloidal gold labeling lap gene monoclonal antibody measuring Listeria monocytogenes kit

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Embodiment 1

[0092] Process conditions:

[0093] The first step is cloning and prokaryotic expression of the L. monocytogenes gene.

[0094] Main equipment:

[0095] Drugs and reagents: strains and plasmids Listeria monocytogenes strain (C5305 strain), prokaryotic expression vector plasmid pET228a are products of TAKARA company. Competent cells Escherichia coli JM109, BL21 recipient bacteria are products of Promega.

[0096] Enzymes and related reagents Ex Taq polymerase, restriction enzymes EcoR 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.

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Abstract

The invention is a colloidal gold labeling lap gene monoclonal antibody measuring Listeria monocytogenes kit, belongs to the technical field of the hygiene analysis and medical test, and is characterized by comprising the following steps: a. primer designing; b. reaction conditions for PCR amplification; c. reaction conditions of agarose gel electrophoresis (AGE); e. applying heat shock conversion method; f. preparing plasmid by SDS cracking process clean up kit; g. induction expression of protein; h. SDS-PAGE electrophoresis; j. cutting the required strips from the SDS-PAGE electrophoresis gel; and k. preparing colloidal gold by using a trisodium citrate reduction method with little improvement. The invention has the beneficial effects of improving the limitation that only single legionella single antigen can be measured. The invention also features pure antibody and high valence and can improve accuracy and sensitivity. Compared with ELISA method and PCR method, the measurement for Listeria monocytogenes in the invention is rapid and convenient by colloid gold-immunochromatography assay.

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.welshmei 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 bacteria of mycosis. Listeria monocytogenes (Usteria monocytogenes, Lm) is a kind of short Gram-positive bacillus, is a kind of zoonotic pathogenic bacteria, can make people and animals suffer from Listeria Mycosis. 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 pig...

Claims

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Application Information

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IPC IPC(8): C12N15/55C12N15/63C12N9/14C07K16/40G01N33/569
Inventor 何成彦贾芙蓉方玲李洪军
Owner 何成彦
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