Porcine circovirus II antibody detection ELISA kit

A porcine circovirus and type 2 antibody technology, applied in the field of vaccine detection, can solve problems such as difficulty in obtaining vaccines and differences in vaccine effects

Inactive Publication Date: 2015-10-28
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, due to differences in vaccine preparation batches, there are differences in the effects of different batches of vaccines
At present, in terms of vaccine evaluation, in addition to the determination of the agar expansion titer after the vaccine is diss

Method used

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  • Porcine circovirus II antibody detection ELISA kit
  • Porcine circovirus II antibody detection ELISA kit
  • Porcine circovirus II antibody detection ELISA kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Preparation and use of porcine circovirus type 2 guinea pig antibody detection ELISA kit

[0016] 1. The preparation method of porcine circovirus type 2 guinea pig antibody detection kit:

[0017] Preparation and purification of PCV2 antigen: Escherichia coli engineering bacteria expressing recombinant porcine circovirus type 2 Cap protein were inoculated with 1% to 2% inoculum in a 2000 ml shake flask for culture, and the culture medium was prepared at a volume of 35%. 1mol / L sodium hydroxide solution to adjust the pH value to 7.0-7.2. Add 10% Kanamycin Sulfate Injection at a ratio of 0.1% of the total volume, incubate at 35-36°C until the OD of the bacterial solution 600nm When the value reaches 0.8-1.0, add sterile 0.5mol / L α-lactose solution to make the final concentration reach 0.03mol / L, and induce expression for 5 hours. Bacteria were collected by centrifugation. The collected bacteria were resuspended with normal saline, ultrasonically destructed, a...

Embodiment 2

[0049] Specificity, sensitivity of the porcine circovirus guinea pig antibody detection ELISA kit of embodiment 2 and carry out comparative test with agar expansion method

[0050] 1. Specificity test: The PCV2 ELISA antibody detection kit developed in this study is for the specific detection of circovirus by the detection kit. Three batches of kits were used to detect porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus respectively. , swine fever, porcine pseudorabies and other 10 kinds of single-factor positive sera (prepared after immunizing guinea pigs with the vaccine) were detected, and the results were negative except for PCV2 positive sera. See Table 1 for details, indicating that the kit has good efficacy. specificity.

[0051] Table 1: Test results of detection specificity of 3 batches of kits

[0052]

[0053] Note: 1 = porcine reproductive and respiratory syndrome positive serum; 2 = swine fever positive serum; 3 = porcine pseudora...

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Abstract

The invention provides a porcine circovirus II antibody detection ELISA kit. The kit comprises an ELISA plate coated with a purified escherichia coli-expressed porcine circovirus II Cap protein as an antigen, positive contrast serum, negative contrast serum, a sample diluent, an enzyme labeling conjugate, a washing solution, a substrate solution A, a substrate solution B and a stopping solution. An indirect ELISA method is adopted, the coated plate is coated with the prokaryotic expressed Cap protein as an antigen and antigen purity is high. The ELISA detection kit prepared from the coated plate with the antigen has high sensitivity, specificity and repeatability.

Description

technical field [0001] The invention belongs to the technical field of vaccine detection, in particular to an ELISA kit for detecting porcine circovirus type 2 antibody. Background technique [0002] Porcine circovirus disease type 2 can cause immunosuppressive disease. The virus can induce mixed infection and secondary infection of various viruses and bacteria, and cause huge economic losses to the pig industry. The research on this disease, especially the research on its vaccine, is attracting more and more attention. Vaccines are the main means of preventing and controlling PCV2. Researchers at home and abroad are researching and developing various vaccines. So far, some vaccines have been successful in clinical trials, some have been put into use, and other candidate vaccines are also under active research. [0003] However, due to differences in vaccine preparation batches, different batches of vaccines have different effects. At present, in terms of vaccine evaluatio...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56983
Inventor 徐保娟荣俊杜元钊范根成郭伟伟刘蕾张恒宫晓朱艳梅王丽萍孙健胡潇
Owner YEBIO BIOENG OF QINGDAO
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