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Sheep echinococcosis ELISA antibody detection kit and application thereof

A technology for antibody detection and amniotic hydatid disease, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of less time-consuming, poor clinical usability, and easy to be affected by the environment, and achieves high sensitivity and specificity. Simple, practical effects

Pending Publication Date: 2020-04-03
北京明日达科技发展有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The indirect hemagglutination test uses cells as the carrier to specifically combine with antigens and antibodies, which has high accuracy, less time-consuming, is easily affected by the environment, and has poor clinical applicability; the polymerase chain reaction has good specificity, sensitivity, and repeatability, but it is not effective for Laboratory hardware requirements are relatively high, and it is difficult to promote in grassroots veterinary laboratories

Method used

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  • Sheep echinococcosis ELISA antibody detection kit and application thereof
  • Sheep echinococcosis ELISA antibody detection kit and application thereof
  • Sheep echinococcosis ELISA antibody detection kit and application thereof

Examples

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Effect test

Embodiment 1

[0026] Embodiment 1, the preparation of sheep echinococcosis ELISA antibody detection kit

[0027] 1. The composition of sheep echinococcosis ELISA antibody detection kit

[0028] Echinococcosis ELISA antibody detection kit consists of Echinococcus granulosus cocktail antigen pre-coated microtiter plate, positive control solution, negative control solution, enzyme marker concentrate, enzyme marker diluent, sample diluent, substrate A solution, substrate B solution, stop solution, washing solution, serum dilution plate composition.

[0029] 2. Preparation of sheep echinococcosis ELISA antibody detection kit

[0030] 1) Preparation of Echinococcus granulosus cocktail antigen

[0031] A. Primer Design

[0032] Primers were designed according to the sequences of EPC-1 (accession number AF481884), EgAgB1 (accession number Z26336.1), EgAgB2 (accession number JN050887.1), EgAgB4 (accession number AY871031.1) in GeneBank:

[0033] EPC-1-F: GGATCC GCCGTACTATGAGTCTTCA

[0034] E...

Embodiment 2

[0121] Embodiment 2, sheep echinococcosis ELISA antibody detection kit detects echinococcosis

[0122] 1. Reagent preparation

[0123] The concentrated washing solution should be returned to room temperature and shaken to dissolve the precipitate, then diluted 25 times with deionized water, mixed well, and the diluted washing solution can be stored at 4°C for 7 days.

[0124] 2. Sample pretreatment

[0125] Whole blood was taken, and after the blood was coagulated, it was centrifuged at 4000rpm for 10min, and the supernatant was collected. The serum can also be coagulated naturally without hemolysis; the serum sample to be tested is diluted in a serum dilution plate at a volume of 1:99 and mixed thoroughly.

[0126] 3. Detection

[0127] 1) Take the Echinococcus granulosus cocktail antigen pre-coated microtiter plate, and add 100 μL of the diluted serum to be tested to the pre-coated microtiter plate. At the same time, 2 wells for negative control and 2 wells for positive ...

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Abstract

The invention discloses a sheep echinococcosis ELISA antibody detection kit and a preparation method and application thereof. In the detection kit, an elisa plate is pre-coated with an echinococcus granulosus cocktail antigen; the echinococcus granulosus cocktail antigen is composed of an EPC1 recombinant antigen, an EgAgB1 recombinant antigen, an EgAgB2 recombinant antigen and an EgAgB4 recombinant antigen. The echinococcus granulosus cocktail antigen is composed of an EPC1 recombinant antigen, an EgAgB1 recombinant antigen, an EgAgB2 recombinant antigen and an EgAgB4 recombinant antigen. Thedetection kit provided by the invention can be used for rapidly detecting the echinococcosis antibody in the sheep serum, is simple to operate, short in consumed time, high in sensitivity and good inspecificity, and is convenient for simultaneously detecting a large number of samples. The sheep echinococcosis ELISA antibody detection kit provided by the invention has important application valuein echinococcosis antibody level detection and epidemiological investigation.

Description

technical field [0001] The invention belongs to the field of enzyme-linked immunosorbent detection, and relates to a sheep echinococcosis ELISA antibody detection kit and a preparation method and application thereof. Background technique [0002] Hydatid disease is a zoonotic disease caused by the larvae of Echinococcus tapeworm parasitizing in humans or animals. my country lists it as a second-class animal disease, and OIE lists it as a B-class animal disease. Echinococcus can parasitize any organ in the host body, oppress the surrounding tissue and cause atrophy and dysfunction, and the rupture of the cyst can cause secondary infection or shock, or even death. There are currently 5 species of Echinococcus tapeworms confirmed, among which the cystic echinococcosis caused by Echinococcus granulosus is concentrated in the large animal husbandry province in Northwest my country, especially in sheep, cattle, and pigs, which poses a threat to public health safety and animal husb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535
CPCG01N33/535G01N2333/43543
Inventor 刘河冰温凯杨柳李蓉蓉马立才
Owner 北京明日达科技发展有限责任公司
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