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A diagnostic kit for detecting paragonimiasis and its preparation method

A diagnostic kit and fluke disease technology, applied to measuring devices, material analysis through observation of the impact on chemical indicators, instruments, etc., can solve problems such as time-consuming, labor-intensive, inaccurate, and achieve no need for special equipment, repeated The effect of good sex and high sensitivity

Inactive Publication Date: 2011-12-14
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a diagnostic kit for detecting Paragonimiasis and a preparation method thereof. The diagnostic kit for detecting Paragonimiasis and its preparation method should solve the problem of detecting Paragonimiasis in the prior art. time-consuming, labor-intensive and inaccurate technical problems

Method used

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  • A diagnostic kit for detecting paragonimiasis and its preparation method
  • A diagnostic kit for detecting paragonimiasis and its preparation method

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Embodiment 1

[0026]The invention provides a diagnostic kit for detecting Paragonimiasis, which contains the polyclonal antibody (rabbit anti-PW-IgG) of rabbit anti-Paragonia wescheri adult worm soluble antigen, and the enzyme-labeled rabbit anti-Paragonia wescheri Polyclonal antibody to adult soluble antigen (HRP-PW-IgG), nitrocellulose film, PBST solution of bovine serum albumin, PBST washing solution, substrate chromogenic solution (A liquid, B liquid, C liquid), positive control Samples, negative control samples, polyethylene reaction plates.

[0027] Further, the PBST solution of bovine serum albumin is composed of bovine serum albumin and PBST washing liquid, and the weight ratio of bovine serum albumin and PBST washing liquid is: 1:99.

[0028] Further, the PBST washing solution is composed of sodium chloride, KH 2 PO 4 、Na 2 HPO 4 . 2 PO 4 、Na 2 HPO 4 , KCl and water, the weight percentage of the sodium chloride in the base liquid is 0.8%, the KH 2 PO The weight percentage ...

Embodiment 2

[0035] The present invention also provides a method for preparing a diagnostic kit for detecting Paragonimiasis, comprising a step of preparing a polyclonal antibody (rabbit anti-PW-IgG) against the adult soluble antigen of Paragonimus wescheri, a step of preparing The step of the polyclonal antibody (HRP-PW-IgG) of the polyclonal antibody (HRP-PW-IgG) of the rabbit anti-Paragonimus wescheri adult worm soluble antigen of enzyme labeling also includes a nitrocellulose film, 1% bovine serum albumin (BSA), Steps for PBST washing solution, substrate chromogenic solution, positive control sample, negative control sample, and polyethylene reaction plate.

[0036] 1. Prepare the polyclonal antibody (rabbit anti-PW-IgG) of the rabbit anti-Paragonia wescheri adult soluble antigen (rabbit anti-PW-IgG) and the polyclonal antibody (HRP-PW-IgG) of the enzyme-labeled rabbit anti-Paragenes wescheri adult soluble antigen The steps are as follows:

[0037] Adults were isolated from the lungs ...

Embodiment 3

[0053] The operation steps of adopting the kit of the present invention to detect Paragonimiasis:

[0054] (1) Dilute rabbit anti-PW-IgG to 200 μg / ml, pipette 1 μl of antibody onto nitrocellulose membrane, and coat at room temperature for 10 minutes.

[0055] (2) Blocking: immerse the coated membrane in 1% bovine serum albumin in PBST solution (BSA) at 37° C. for 15 minutes.

[0056] (3) Washing: Wash the sealed membrane 3 times with washing solution, and dry it for 3 minutes each time, which is the "quick diagnosis membrane".

[0057](4) Antigen to be tested: Cut out the "quick diagnosis membrane" according to the grid, put the glossy side up, put it into the well of a 96-well polyethylene reaction plate, add 100 μl of the serum to be tested, and set blank (PBS), negative and Positive sample control, 37 ℃ 1h.

[0058] (5) Washing is the same as (3)

[0059] (6) Enzyme-labeled HRP-PW-IgG: Dilute the enzyme-labeled antibody 1:50, add 100 μl to each well, and incubate at 37° ...

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Abstract

The invention provides a diagnostic kit for detecting Paragonimiasis, which contains a rabbit polyclonal antibody against the soluble antigen of Paragonia wescheri adult worm, an enzyme-labeled rabbit polyclonal antibody against the soluble antigen of Paragonimus westermani adult worm, Nitrocellulose membrane, bovine serum albumin, PBST washing solution, substrate chromogenic solution, positive control sample, negative control sample and polyethylene reaction plate. The invention also provides a preparation method of a diagnostic kit for detecting paragonimiasis. The invention adopts the double-antibody sandwich method Dot-ELISA to detect the circulating antigen of Paragonimus westermani, has high sensitivity and strong specificity, and can realize rapid diagnosis and curative effect assessment of Paragonimiasis.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and a preparation method thereof, in particular to a diagnostic kit for detecting Paragonimiasis and a preparation method thereof. Background technique: [0002] Paragonimiasis (paragonimiasis) is a food-borne parasitic disease that endangers human health. It has been found in China for 125 years. The first case was found in the lungs of Taiwanese Portuguese sailors by Ringer. There are more than 50 species of worms reported in the world at present, among which 28 species have been reported in various places in China, and reports of this disease have been found successively in more than 20 provinces (regions) in my country. In recent years, with the deepening of reform and opening up, due to the flow of population and the change of people's diet structure, paragonimiasis patients have a tendency to spread from rural areas to cities, and paragonimiasis has become ...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/531G01N21/78
Inventor 陈韶红陈家旭李浩周晓农张永年郭俭
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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