ELISA kit for detecting porcine circovirus antibody II

A porcine circovirus, type 2 antibody technology, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as high background and false positives, and achieve short detection time, near-sensitivity, and simple operation process. Effect

Active Publication Date: 2010-01-20
北京金诺百泰生物技术有限公司
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AI Technical Summary

Problems solved by technology

The ELISA method currently used to check PCV2 antibodies is mainly an indirect method. The disadvantage of this method is that the high concentration of non-specific antibodies contained in the serum can be directly adsorbed on the solid phase carrier, and sometimes also on the surface of the coated antigen. , resulting in high background or possible false positive results

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  • ELISA kit for detecting porcine circovirus antibody II
  • ELISA kit for detecting porcine circovirus antibody II
  • ELISA kit for detecting porcine circovirus antibody II

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Embodiment Construction

[0026] Preparation of coating antigen:

[0027] According to the gene sequence of PCV2 in GenBank (Accession No.: AY943819), two primers were designed, and the above primers were: GC GAATTC ATGGGCATCTTCAACACCCGCCTCTC; downstream primer is: GC GTC GACTTACTTAGGGTTAAGTGGGGGGTC. Amplify the Cap gene without nuclear localization signal from the porcine PCV2 genome by PCR, clone it into the pET32-a(+) prokaryotic expression vector (purchased from Novagen, product number: 69015-3), and construct pET32a -Cap recombinant expression vector. Transformed into Escherichia coli BL21(DE3), and expressed the recombinant Cap fusion protein with reference to Novagen's pET Syetem Manual, and the protein was expressed in a soluble manner. Purify the expressed fusion protein with affinity chromatography (Novagen His-BindKit, product number: 70239-3), see figure 1 .

[0028] Preparation of PCV2 antibody detection plate:

[0029] Dilute the purified Cap fusion protein with 0.05M carbonate bu...

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Abstract

The invention relates to an ELISA kit for detecting porcine circovirus antibodies II, wherein, the kit is developed according to the double antigen sandwich ELISA principle. The ELISA kit comprises a prepacked ELISA antigen plate bar, a cleaning solution, 100 times concentrated enzyme labeled antigen, an enzyme labeled dilution, a zymolyte coloration, a stopping solution, standard PCV2 negative sera, and standard PCV2 positive sera. The kit replaces enzyme labeled anti-antibodies with the enzyme labeled antigen. The enzyme labeled antigen can not combine with antibodies absorbed with ELSA plate without specificity. Meanwhile, the serum specimen to be detected does not need to be pre-diluted, and the serum specimen can be directly mixed with the enzyme labeled antigen with working concentration to be directly used for determination. The ELISA kit has the advantages of simple operation and shorter detecting time.

Description

Technical field: [0001] The invention relates to an ELISA kit for detecting porcine circovirus type 2 (PCV2) antibody, belonging to the technical field of veterinary biology. It is used for the antibody detection of porcine circovirus 2, so as to judge the infection situation of porcine circovirus type 2 in pig herds and monitor the prevalence of the disease and the immune effect. Background technique: [0002] Porcine circovirus type 2 (porcine circovirus 2, PCV2) is closely related to many diseases in pigs, such as post weaning multisystem wasting syndrome (PMWS), porcine dermatitis and nephropathy syndrome , PDNS), sow reproductive disorder, proliferative enteritis, piglet congenital tremor, porcine respiratory disease syndrome, etc., we collectively refer to various diseases caused by PCV2 as PCVD (porcinecircovirus disease, PCVD), among many PCVD, PMWS Most commonly, PMWS causes progressive wasting, diarrhoea, respiratory symptoms, pale or jaundiced skin, and reduced p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/535G01N33/52C12N15/70C12P21/02C12R1/19
Inventor 余兴龙葛猛李润成蒋大良罗维
Owner 北京金诺百泰生物技术有限公司
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