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Indirect ELISA diagnostic reagent kit for pig B type circular virus antibody

A diagnostic kit, circovirus technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as time-consuming and complicated test operations, and achieve simple and fast operation, broad market prospects, high specificity and sensitivity. Effect

Inactive Publication Date: 2005-03-02
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above techniques and methods can detect PCV2 and its antibody levels, and have achieved certain results in practice, they all have complex test operations, time-consuming, specific professional skills and equipment, etc., and are often limited to laboratories. Difficult to popularize and promote at the grassroots level

Method used

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  • Indirect ELISA diagnostic reagent kit for pig B type circular virus antibody
  • Indirect ELISA diagnostic reagent kit for pig B type circular virus antibody

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0020] (1) Preparation of PCV2 recombinant dCap protein

[0021] Design specific primers according to the PCV2 Cap gene sequence (upstream primer: 5'-GC GGATCC AATGGCATCT TCAACAC-3'; downstream primer: 5'-CCG CTCGA GTTAAGGGTTAAGTGGG-3'), utilize PCR technology to amplify the capsid protein gene sequence (cap gene 3' end 579bp fragment) of porcine PCV2 genome from the porcine PCV2 genome (see Figure 3), insert it into the prokaryotic expression vector pGEX The pGEX-PCV2-dCap expression vector was constructed downstream of the glutathione-S-transferase gene (GST) of -4T-1, and transformed into Escherichia coli BL21, and after being induced and expressed by 0.1mM IPTG for 5 hours, the bacteria were lysed by ultrasonic waves, and used The expressed fusion protein (GST-dCap) was purified by a commercial GST affinity chromatography column, and then the GST tag was removed by enzymatic hydrolysis with thrombin to obtain the recombinant nuclear localization signal capsid protein of...

Embodiment

[0048] Implementation Example PCV2 Antibody Indirect ELISA Diagnostic Kit

[0049] Principle see figure 1 , prepare recombinant PCV2 dCap protein according to (1) method step in the embodiment, prepare PCV2 antibody detection plate according to (2) method step in the embodiment, prepare PCV2 antibody detection kit enzyme by (3)(4) method step in the embodiment For the working solution of the conjugate, prepare the sample diluent, washing solution, and stop solution according to the method step (5) in the embodiment, and prepare the positive control and negative control of the antibody detection kit according to the method step (6) in the embodiment, according to the method step in the embodiment ( 7) Method steps Prepare color developing solution A and color developing solution B, and finally, assemble various components into a PCV2 antibody detection kit.

[0050] When using the PCV2 antibody indirect ELISA diagnostic kit to detect porcine PCV2 antibody or antibody level, th...

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PUM

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Abstract

The invention discloses an indirect ELISA diagnosing reagent box for pig II type circular virus (PCV2) antibody. There arranged with antibody detecting board in the reagent box, enzyme combination liquid, positive comparison, negative comparison, sample diluter, 10X condensed detergent, developing liquid A and B, and stopping liquid, the detecting board of the reagent box is removable 96-apertures enzyme labeling board enclosed with PCV2 reconstructed enucleation localizing signal capsid protein (dCap), the enzyme combination liquid is goat-anti-pig IgG multi-clone antibody labeled with HRP, the positive comparison is pig PCV 2 standard positive serum, the negative comparison is pig standard negative serum. The sensitivity of the invention is high, simple, and easy to be wide applied.

Description

technical field [0001] The invention relates to a rapid diagnosis method and equipment for a virus antibody of pigs, in particular to an indirect ELISA diagnostic kit for antibodies to porcine type II circovirus (PCV2). Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) is the smallest animal virus found so far. The diameter of the virus particle is about 17nm. It is a covalently closed, circular, single-stranded DNA virus. PCV has two genotypes: PCV1 and PCV2. PCV1 is non-pathogenic, but widely exists in pigs and pig-derived cell lines. PCV2 is pathogenic to pigs and mainly causes post-weaning multisystemic wasting syndrome (PMWS) in piglets. PMWS is a new infectious disease, first discovered in Canada in 1991, mainly occurs in weaned piglets aged 5-12 weeks, manifested as progressive emaciation, dyspnea, pale skin, diarrhea, and jaundice. Subsequently, the United States, the United Kingdom, Germany, France, Ireland, the Czech Republic, Spain, China...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/547
Inventor 周继勇商绍彬申会刚陈庆新
Owner ZHEJIANG UNIV
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