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Staphylococcal enterotoxin A chemiluminiscence enzyme-linked immunoassay detection kit

A staphylococcal enteric and chemiluminescent enzyme technology, which is applied in the direction of chemiluminescence/bioluminescence, analysis of materials through chemical reactions, and analysis of materials, can solve the problems of many interference factors, expensive, time-consuming, etc., and achieve repeatability Good, strong specificity, high sensitivity effect

Inactive Publication Date: 2013-02-13
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of immunoagar diffusion method and reverse indirect hemagglutination method are time-consuming, many interference factors affecting the analysis, and poor specificity.
The immunochip method is expensive, operators need to undergo professional training, and there are many interference factors that affect the analysis.
Enzyme-linked immunosorbent assay is sometimes not sensitive enough to detect

Method used

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  • Staphylococcal enterotoxin A chemiluminiscence enzyme-linked immunoassay detection kit
  • Staphylococcal enterotoxin A chemiluminiscence enzyme-linked immunoassay detection kit
  • Staphylococcal enterotoxin A chemiluminiscence enzyme-linked immunoassay detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Preparation of Staphylococcus aureus Enterotoxin A Monoclonal Antibody and Screening of Coating Antibody and Detection Antibody Pairing

[0046] (1) Preparation of monoclonal antibody against Staphylococcus aureus enterotoxin A: The purified natural Staphylococcus aureus enterotoxin A antigen standard was provided by the Institute of Microbial Epidemiology, Academy of Military Medical Sciences. Eight-week-old BALB / c mice were used as immunized animals. Staphylococcus aureus enterotoxin A was used as the immunogen, and the first immunization dose was 50 μg / mL. The immunogen was dissolved in normal saline and an equal volume of Freund’s complete adjuvant. Emulsifier, multi-point subcutaneous injection. The second immunization was carried out after an interval of 3 weeks. The dose and route were the same as the first immunization, and the adjuvant was Freund's incomplete adjuvant. After 2-3 weeks, the third immunization was carried out, the dose was the same as...

Embodiment 2

[0053] Example 2: Establishment of Chemiluminescent ELISA Method

[0054] (1) Selection of optimal coating antibody concentration: Coating antibody is coated with chemiluminescent enzyme at serial dilutions of 1.25 μg / mL, 2.5 μg / mL, 5 μg / mL, 10 μg / mL, 20 μg / mL and 40 μg / mL Connect plates, 100 μL / well, incubate overnight at 4°C, wash the plate 3 times with washing solution; seal with 250 μL / well blocking solution, place at room temperature for 1 hour, wash the plate 3 times; add 0.01ng / mL Staphylococcus aureus enterotoxin A Standard antigen, 100 μL / well, at the same time each coating antibody concentration was used as a blank control well, incubated at 37°C for 1 hour, and washed 3 times; respectively added 1:4000 diluted anti-Staphylococcus aureus enterotoxin A detection antibody, 37 Incubate at ℃ for 1 hour, wash the plate 5 times; add 100 μL / well of chemiluminescence solution, and measure the luminescence value. According to the ratio of the measured different coating antib...

Embodiment 3

[0064] Example 3: Application of chemiluminescent ELISA detection kit for detection of Staphylococcus aureus enterotoxin A in different matrices

[0065] This example is used to evaluate the accuracy and practicability of the staphylococcal enterotoxin A chemiluminescent ELISA detection kit of the present invention in detecting staphylococcus aureus enterotoxin A in different matrices.

[0066] Dilute the standard Staphylococcus aureus enterotoxin A with environmental matrix river water, food matrix milk, body fluid matrix human serum or human urine to different concentrations, detect the content of Staphylococcus aureus enterotoxin A by chemiluminescent enzyme immunoassay, and calculate The ratio of the measured concentration to the actual concentration added is the recovery rate.

[0067] (1) Coat the chemiluminescence enzyme-linked plate with the coating antibody: Dilute the coating antibody to 2.5 μg / mL with the coating solution and add to the chemiluminescence enzyme-link...

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Abstract

The invention discloses a staphylococcus aureus enterotoxin A chemiluminiscence enzyme-linked immunoassay detection kit. The kit contains a monoclonal antibody resisting staphylococcus aureus enterotoxin A, horseradish peroxidase marked detection antibody solution resisting staphylococcus aureus enterotoxin A, staphylococcus aureus enterotoxin A serial standard solution, wrapping solution, washing solution, closing solution, chemiluminiscence solution and a chemiluminiscence enzyme-linked plate. The staphylococcus aureus enterotoxin A chemiluminiscence enzyme-linked immunoassay detection kit has the advantages of being strong in specificity, high in sensitivity and good in repeatability and is suitable for monitoring diseases and detecting staphylococcus aureus enterotoxin A in food and blood in rapid quantitative mode.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and relates to a superantigen detection kit, in particular to a staphylococcus aureus enterotoxin A chemiluminescent enzyme-linked immunoassay detection kit, which utilizes a double-antibody sandwich chemiluminescent enzyme Linked immunoassay to quantitatively detect the content of Staphylococcus aureus enterotoxin A in the environment, food and body fluids such as river water, milk, urine and blood. Background technique [0002] Staphylococcus aureus is an important pathogenic bacterium of zoonosis, which widely exists in air, water, dust, and human and animal excreta. According to the U.S. Centers for Disease Control and Prevention report, food poisoning caused by Staphylococcus aureus is second only to Escherichia coli, accounting for 33% of bacterial food poisoning. Canada reported higher rates, accounting for 45% of bacterial food poisoning cases. There are also many such poisoning...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/76
Inventor 张春梅李永明宋朝君徐竹蔚李琦黄业青刘飞刘志佳杨琨陈丽华张赟方亮孙元杰易静周幸春马樱刘蓓张宇丝刘蓉蓉金伯泉
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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