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Cytology detection kit for discriminating cholera toxin and detection method

A technology for detecting kits and cholera toxins, which is applied in biochemical equipment and methods, and microbial determination/inspection, etc., can solve problems such as low sensitivity, long detection cycle, and inability to identify cholera toxin activity, and achieve enhanced monitoring and control, Improve the effect and efficiency of the effect

Inactive Publication Date: 2012-07-04
ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, traditional CT detection methods mainly rely on animal experiments, traditional cytology, immunology, molecular biology and biosensor technologies, but these methods have a long detection cycle, low sensitivity, and cannot effectively identify the activity of cholera toxin

Method used

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  • Cytology detection kit for discriminating cholera toxin and detection method
  • Cytology detection kit for discriminating cholera toxin and detection method
  • Cytology detection kit for discriminating cholera toxin and detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0022] 1. Materials:

[0023] Y1 adrenocortical cells (ATCC: CCL-79), F-12K medium, and horse serum were purchased from ATCC; fetal bovine serum was purchased from Hyclone; cholera toxin was purchased from List biological laboratories; polylysine, bovine serum Albumin and anti-cholera toxin neutralizing antibody were purchased from Sigma Company; 96-E-plate was purchased from Swiss Roche Company. The real-time cell analysis system was purchased from Roche, Switzerland.

[0024] 2. Detection method:

[0025] 2.1 Coating and cell plating of 96-E-plate

[0026] Dilute poly-lysine with phosphate buffer (pH7.2) according to 1:4 (v / v) to prepare coating solution, add 100 μl volume of coating solution to each well to coat 96-E-plate, and let it stand at room temperature. Set aside for 10 minutes, remove the coating solution and wash twice with phosphate buffered saline, and set aside.

[0027] Add horse serum and fetal bovine serum to F-12K medium according to 15% and 2.5%, respe...

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Abstract

The invention provides a cytology detection kit for discriminating cholera toxin and a detection method. The kit mainly comprises Y1 adrenocortical cells, anti-cholera toxin neutralizing antibody, polylysine, 96-E-plate and cell culture medium for the adrenocortical cells. The invention provides the cytology detection kit for discriminating the cholera toxin and the detection method, and with respect to the specific cellular reaction of the cholera toxin, a system capable of dynamically and sensitively detecting the cholera toxin in real time is established, therefore, the cholera toxin can be fast and accurately identified with high specificity, strains producing toxin can be effectively discriminated from strains producing no toxin, the monitoring and control of the strains producing toxin are strengthened, and effects and benefits of prevention can be greatly improved. The invention has important significance for preventing and controlling cholera.

Description

(1) Technical field [0001] The invention relates to a cytology detection kit for discriminating cholera toxin and a method for detecting cholera toxin by using the kit. (2) Background technology [0002] In the prevention and control of Vibrio cholerae epidemics, whether the bacteria produce CT is an extremely important detection index. Almost all the epidemic strains of Vibrio cholerae were CT-producing strains, and none of the non-epidemic strains produced CT. CT is encoded by the ctxAB gene, which is a toxic protein with ADP-ribosyltransferase activity secreted by Vibrio cholerae. This gene is located on the chromosome CTX genetic unit, and its 415kb core region contains at least 5 genes (cep, orfU, ace, zot, ctxAB), carried by lysogenic phage CTXΦ, can be transferred horizontally among Vibrio cholerae. CT toxin consists of five B subunits and one A subunit. The B subunit acts on the intestinal mucosal epithelial cell-specific receptor GM1 (Gangliosidoses, ganglioside m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/02
Inventor 金大智罗芸张政
Owner ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION
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