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A kind of trichinella spiralis competition ELISA antibody detection kit and its detection method

A detection kit and antibody detection technology, applied in the field of serological immunodetection, to achieve the effect of strong practicability, high specificity and strong specificity

Active Publication Date: 2022-05-31
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional solid-phase competitive ELISA is to coat the antigen on a 96-well microwell plate, and the serum to be tested competes with the monoclonal antibody to bind to the antigen on the bottom of the microwell plate, and then uses the HRP-labeled secondary antibody against the monoclonal antibody for signal detection, but due to Monoclonal antibodies are limited in species origin (mainly mice and rabbits), and can only be used in host detection projects that are not derived from monoclonal antibody species

Method used

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  • A kind of trichinella spiralis competition ELISA antibody detection kit and its detection method
  • A kind of trichinella spiralis competition ELISA antibody detection kit and its detection method
  • A kind of trichinella spiralis competition ELISA antibody detection kit and its detection method

Examples

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Embodiment 1

[0066] Example 1. Trichinella spiralis competition ELISA antibody detection kit.

[0067] A Trichinella spiralis competition ELISA antibody detection kit described in this example contains: a 96-well reaction plate coated with Trichinella spiralis Ts-WN10 recombinant antigen, and a biotin-labeled monoclonal antibody diluted to 2 μg / mL with 0.9% NaCL solution Cloning antibody Ts-WN10-1H9 solution, horseradish catalase HRP-labeled avidin solution, washing solution, TMB chromogenic substrate, stop solution, and negative quality control; wherein, the negative quality control is a concentration of 1 μg / mL biotin-labeled monoclonal antibody Ts-WN10-1H9 solution; the monoclonal antibody Ts-WN10-1H9 is prepared by hybridoma cell line WN10-1H9, and the hybridoma cell line WN10-1H9 The microbial preservation number is CGMCC No.18316. The cell line was preserved in the General Microbiology Center of the China Microbiological Culture Collection Management Committee on August 15, 2019. The...

Embodiment 2

[0087] Example 2, the detection method of Trichinella spiralis competition ELISA antibody detection kit.

[0088] 1) Detection method of serum samples:

[0089] Take 50 μL of biotin-labeled monoclonal antibody Ts-WN10-1H9 solution and 50 μL of the serum to be tested, mix them in EP tubes, add them to the 96-well reaction plate pre-coated with Ts-WN10 recombinant antigen, and compete for reaction at 37°C 1h, set up a negative quality control control (1μg / mL, 100μL) in the plate;

[0090] After discarding the reaction solution, add 300 μL PBST washing solution to each well, wash 3 times, 1 min each time;

[0091] Dilute HRP-labeled avidin with 1% BSA-containing PBST solution at 1:400, add 100 μL to each reaction well, and incubate at 37°C for 30 minutes;

[0092] Discard the reaction solution, add 300 μL PBST washing solution to each well, and wash 3 times, each time for 1 min;

[0093] Add 100 μL of TMB substrate to each well, develop color at 37°C for 8 min, then add 50 μL ...

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Abstract

A trichinella spiralis competition ELISA antibody detection kit and a detection method thereof belong to the technical field of serological immunological detection. In order to detect multi-host Trichinella infection more stably and accurately, the invention provides a Trichinella competitive ELISA antibody detection kit, which contains: a 96-well reaction plate whose bottom plate is coated with Trichinella Ts-WN10 recombinant antigen, Biotin-labeled monoclonal antibody Ts-WN10-1H9 solution, horseradish catalase HRP-labeled avidin solution, washing solution, TMB chromogenic substrate, stop solution, and negative quality control; wherein, the negative The quality control is a solution of biotin-labeled monoclonal antibody Ts‑WN10‑1H9 at a concentration of 1 μg / mL. The kit prepared by the invention has the characteristics of strong specificity, high sensitivity, etc., and can be used for antibody detection of multi-host trichinellosis such as pigs / rats / humans.

Description

technical field [0001] The invention belongs to the technical field of serological immune detection, and in particular relates to a trichinella competition ELISA antibody detection kit and a detection method thereof. Background technique [0002] Trichinellosis is a very serious zoonotic disease. It not only causes huge economic losses to animal husbandry production, but also poses a huge threat to human health. People or animals mainly contain trichinosis through raw or semi-raw food. Caterpillar meat (mainly pork) and disease. [0003] For the inspection of trichinellosis in slaughtered animals, the inspection methods of the regulations of the International Organization for Animal Health (OIE) are microscope inspection and sample digestion method, and these two methods are also being used in my country at present. However, the above two methods have certain disadvantages. Microscopic examination is time-consuming and laborious and has poor sensitivity. The sensitivity can...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/577G01N33/535
CPCG01N33/68G01N33/543G01N33/577G01N33/535G01N2333/4353
Inventor 刘晓雷刘明远白雪杨勇王学林丁静刘琰
Owner JILIN UNIV
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