A kit for detecting circulating antigens of Agkistrodon acutina using specific polyclonal antibodies

A technology of Agkistrodon acutus and Ligulata, applied in the field of biological detection, to achieve rapid diagnosis, considerable economic and social benefits, and high practical value

Inactive Publication Date: 2011-12-21
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

Coupled with the expansion of the endemic area of ​​lingualworm in the past 20 years, the discovery of new pathogenic species and the increase of fatal cases, the diagnosis of lingualworm, a rare parasitic disease, poses new challenges

Method used

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  • A kit for detecting circulating antigens of Agkistrodon acutina using specific polyclonal antibodies
  • A kit for detecting circulating antigens of Agkistrodon acutina using specific polyclonal antibodies

Examples

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Embodiment 1

[0022] Example 1 A kit for detecting circulating antigens of Agkistrodon acutina using specific polyantibody

[0023] The invention provides a kit for detecting circulating antigens of Agkistrodon acutinosa larvae using specific polyantibodies, including rabbit polyclonal antibody (rabbit anti-Aag-IgG), horseradish Oxidase-labeled polyclonal antibody against Agkistrodon acutinae (HRP-Aag-IgG), nitrocellulose membrane, 1% bovine serum albumin (BSA) in PBST solution, PBST washing solution, substrate color development solution, positive and negative control samples, and 96-well polyethylene reaction plates.

[0024] Further, the 1% bovine serum albumin in PBST solution is composed of bovine serum albumin and PBST washing solution, and the weight ratio of the bovine serum albumin and PBST washing solution is 1:99.

[0025] Further, the PBST washing solution is composed of sodium chloride, KH 2 PO 4 、Na 2 HPO4 . 2 PO 4 、Na 2 HPO 4 , KCl and water, the weight percentage of t...

Embodiment 2

[0030] The present invention also provides a preparation method of a kit for detecting circulating antigens of Agkistrodon akistrodon larvae by using specific polyclonal antibodies, including a step of preparing a soluble antigen against Agkistrodon larvae, and a step of preparing a rabbit antibody -Aag-IgG step, an enzyme-labeled antibody (HRP-Aag-IgG) step, also includes a nitrocellulose membrane, 1% bovine serum albumin (BSA) in PBST solution, PBST wash in the kit Solution, substrate chromogenic solution, positive control sample, negative control sample, and polyethylene reaction plate.

[0031] Further, in a step of preparing the soluble antigen of Agkistrodon larvae, the larvae are isolated from the abdomen of mice infected with Agkistrodon larvae eggs for 3-5 months, and the larval soluble antigen is prepared.

[0032] Further, in a step of preparing rabbit anti-Aag-IgG, mix the soluble antigen (AagA) of the larvae of Agkistrodon akistrodon and the complete Freund's adju...

Embodiment 3

[0053] A kind of operation step of the kit that adopts specific polyclonal antibody to detect the circulating antigen of Agkistrodon acutina of the present invention is as follows (principle is as follows: figure 1 shown):

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

[0055] b) Blocking: immerse the coated membrane in a PBST solution of 1% bovine serum albumin (BSA). 37°C, 15 minutes.

[0056] c) 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] d) 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 in each well, and set a blank (PBS), negative compared with positive samples. 37°C for 1h.

[0058] e) Washin...

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Abstract

The invention provides a kit for detecting circulating antigens of Agkistrodon acutinae using specific polyantibody, comprising rabbit anti-Aag-IgG, enzyme-labeled antibody (HRP-Aag-IgG), nitrocellulose film, 1 % bovine serum albumin in PBST solution, PBST washing solution, substrate chromogenic solution, positive and negative control sample composition and polyethylene reaction plate. The invention also provides a preparation method of a kit for detecting circulating antigens of Agkistrodon acutina by using specific polyantibody. The invention adopts the polyclonal antibody to measure the circulating antigen, which is beneficial to making early diagnosis and helping to judge the prognosis. When the parasites in the body die, the circulating antigens disappear quickly, which can be used for efficacy assessment. The kit of the invention has high sensitivity and strong specificity, and can realize rapid diagnosis, drug screening and curative effect assessment of Agkistrodon acutinae ligamentiworm disease.

Description

Technical field: [0001] The invention belongs to the field of biological detection, and in particular relates to a kit and a preparation method thereof, in particular to a kit for detecting circulating antigens of Agkistrodon acutina using specific polyclonal antibodies. Background technique: [0002] Ligellasis is a zoonotic disease of zoonotic origin. There are about 118 known species of tongue worms in the world. There are 6 genera and 10 species of tongue worms that parasitize humans, namely Linguatula serrata, Armillifer armillatus, Agkistrodon A. aggkistrodontis, A. moniliformis, A. grandis, Raillietiella hemidactyli, Leiperia cincinnalis ), Porocephalus crotali, P. Sebekia and P. taiwana etc. The tongue worms that parasitize the human body are recorded in our country include Serratoglossus, Akikikiki Agkistrodon, and Formosanus. In recent years, in Taiwan, Zhejiang, and Guangxi of my country, there have been reports of many cases of Agkistrodon agaricus tongue worm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/544G01N21/78C07K16/18C07K16/06
Inventor 陈韶红陈家旭张永年李浩周晓农
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
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