Enzyme-linked immuno sorbent assay (ELISA) kit created on the basis of hog cholera virus recombinant protein nopaline synthase (NS2)

A recombinant protein and swine fever virus technology, applied in the biological field, can solve the problems of less research on diagnostic methods, and achieve the effect of easy results and low cost

Inactive Publication Date: 2012-12-26
ZHENGZHOU HOUYI PHARMA
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AI Technical Summary

Problems solved by technology

In recent years, most of the domestic and foreign studies on CSF have focused on the research and development of diagnostic methods based on the structural proteins E1 and E2 of CSF, but less research on diagnostic methods based on the non-structural protein NS2 of CSFV

Method used

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Embodiment Construction

[0030] The concrete composition of kit of the present invention is as follows:

[0031] 1. Materials

[0032] 1.1 Reference serum standard Positive serum is 20-25 days after piglets have been immunized with the attenuated CSF vaccine C strain, and the blood is collected to separate the serum after 2 weeks. The HI titer is 1:1024-1:2048 as determined by the HI test. After packaging- Store at 80°C. Negative sera were swine sera that had never been vaccinated and had antibody levels close to the average value of negative sera.

[0033] 1.2 The ELISA reaction plate was produced by Nunc Company in Denmark, and the anti-pig IgG enzyme-labeled antibody was purchased from Sigma Company.

[0034] 1.3 The expression vector PEGFP-C1 was purchased from TAKARA Company.

[0035] 1.4 Experimental animal piglets are all products of Henan Piggy Farm Company.

[0036] 2. Antigen preparation

[0037] The preparation of the coated antigen refers to the genome sequence of the lanate attenuate...

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Abstract

The invention relates to an enzyme-linked immuno sorbent assay (ELISA) kit created on the basis of hog cholera virus recombinant protein nopaline synthase (NS2). The ELISA kit comprises an elisa plate wrapped by the hog cholera virus recombinant protein NS2, a ten-time concentration detergent, a serum diluent, a standard serum, an anti-swine Intravenous gamma globulin (IgG) enzyme-labeled antibody, an nzyme-labeled antibody diluent, a sealing agent, a color developing agent A, a color developing agent B and a stop solution. The ELISA kit can detect hog cholera virus antibodies. The method created by the ELISA kit has specificity, sensitivity and operability. In a sensitivity test, the coincidence rate between the method of the ELISA kit and an immunofluorescence technical method is 82%, and the method of the ELISA kit is more sensitive than the immunofluorescence technical method. The detection rate of other etiological agents through the method of the ELISA kit is zero. Compared with the hemagglutination inhibition (HI) method, the method of the ELISA kit has the advantages of being fast and low in cost, and having easily judging results, and can carry out detection work after hog cholera lapinized virus vaccine immunity.

Description

technical field [0001] The invention relates to an ELISA kit established with the recombinant protein NS2 of swine fever virus, belonging to the field of biotechnology. Background technique [0002] Classical swine fever (CSF) is a highly contagious and fatal infectious disease of pigs caused by classical swine fever virus (CSFV). Clinically, it is characterized by persistent high fever and massive bleeding on the skin and mucous membranes. Points are the main features. CSF is highly contagious, mainly transmitted through the respiratory tract and digestive tract, regardless of season, the transmission speed is fast, the morbidity and mortality are high, and the epidemic is widespread. It has caused huge economic losses to the pig industry and is a serious hazard to my country's pig industry. one of the most important diseases. Our country classifies it as a first-class infectious disease. Therefore, it is of great significance to strengthen the research on CSFV diagnostic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
Inventor 吴红云范金红徐进
Owner ZHENGZHOU HOUYI PHARMA
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