Kit for detecting pig pseudorabies and application thereof

A technology of porcine pseudorabies and porcine pseudorabies virus, applied in the field of kits for detecting porcine pseudorabies antibodies, can solve the problems of low specificity, prone to false positives, low sensitivity, etc., and achieves low detection cost and is not easy to be false Positive or false positive error judgment, high sensitivity effect

Active Publication Date: 2013-10-16
HENAN FENGHUA BREEDING SHARE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, indirect gE-ELISA methods are generally used to detect porcine pseudorabies virus gE protein antibodies. The specificity of these methods is not very strong, the sensitivity is not high, and false positives and false negatives are prone to occur, which will inevitably cause misjudgment and incur unnecessary losses

Method used

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  • Kit for detecting pig pseudorabies and application thereof
  • Kit for detecting pig pseudorabies and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The kit for detecting porcine pseudorabies antibody provided in this example includes an enzyme-linked plate coated with porcine pseudorabies virus gE antigen, rabbit anti-pig IgG antibody labeled with horseradish peroxidase, rapid coating solution, and rapid blocking solution , sample diluent, TMB chromogenic solution, stop solution, 20 times concentrated washing solution, negative control, positive control.

[0048] The fast coating solution is NaOH with a concentration of 0.1 mol / L and a pH of 13; the fast blocking solution is KOH with a concentration of 0.1 mol / L and a pH of 13.

[0049] The sample dilution is obtained by adding casein and NaN in PBS at a concentration of 0.01M and a pH of 7.2 3 Prepared with casein and NaN in the sample diluent 3 The mass fractions were 1% and 0.001%, respectively.

[0050] The TMB concentration in the described TMB chromogen is 0.3g / L; The stop solution is that the concentration is 2M H 2 SO 4 Solution; Among them, TMB chromog...

Embodiment 2

[0069] The present embodiment provides the method for detecting the pseudorabies antibody with the kit for detecting the pseudorabies antibody, comprising the following steps:

[0070] (1) Test sample preparation: After blood collection, centrifuge at 3000 rpm for 5 minutes, and take the supernatant to obtain serum;

[0071] (2) Equilibration: Take the kit out of the refrigerated environment and equilibrate at room temperature for 30 minutes before use;

[0072] (3) With lotion: dilute 20 times concentrated lotion with double distilled water 20 times for later use;

[0073] (4) Setting: Set wells A1-A6 in the microtiter plate, where no sample diluent is added to wells A1 and A2, 100 μL positive control serum is added to wells A3 and A4, and 100 μL negative control is added to wells A5 and A6 Serum, that is, wells A1 and A2 are blank controls, wells A3 and A4 are positive controls, and wells A5 and A6 are negative controls;

[0074] (5) Adding samples: Add 100 μL of detection...

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Abstract

The invention discloses a kit for detecting pig pseudorabies. The kit comprises an assay plate coated with pig pseudorabies virus gE antigen, a rabbit anti-pig IgG antibody marked by HRP (horseradish peroxidase), fast coating buffer, fast blocking buffer, sample diluents, TMB substrate, stop buffer, 20 times wash solution, negative control, and a positive control. Through adopting pig pseudorabies virus specificity gE antigen with high purity and high activity expressed through gene engineering to coat the assay plate, and through adopting HRP-conjugate of rabbit anti-pig IgG monoclonal antibody containing HRP, the kit for detecting pig pseudorabies, provided by the invention, has the advantages of strong specificity, high sensibility, less susceptibility to misjudge of false positive or false negative and the like; moreover, the kit is simple in structure, low in detection cost, convenient and fast in operation and strong in timeliness, has a wide market prospect, and can create larger social benefits.

Description

technical field [0001] The invention relates to the field of animal epidemic detection, in particular to a kit for detecting porcine pseudorabies antibodies. Background technique [0002] Pseudorabies (Pseudorabies, PR) is an acute infectious disease caused by pseudorabies virus (Pseudorabies virus, PRV), a variety of domestic animals and wild animals can be infected, and pigs are the most common infection, causing great harm to the pig industry Economic loss, vaccination is the most effective way to prevent this disease. The current pseudorabies vaccines on the market are gE gene-deficient vaccines, and the vaccinated pigs do not contain porcine pseudorabies virus gE protein antibodies in their serum. If the gE protein antibody is detected in the pig serum, it indicates that the pig has been infected by the wild virus of pseudorabies virus, and the infected pigs can be eliminated to eradicate pseudorabies. [0003] At present, indirect gE-ELISA methods are generally used ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543
Inventor 魏冬青窦文茂
Owner HENAN FENGHUA BREEDING SHARE
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