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Seneca virus blocking ELISA (enzyme-linked immuno sorbent assay) antibody detection kit

An antibody detection and kit technology, which is applied in the field of Seneca virus antibody detection kits, can solve the problems of complicated operation process and result calculation, high professional technical level requirements, unsuitable pig herd infectivity detection, etc., and achieves good results. The effect of reaction sensitivity, simple and fast preparation process, and loose technical requirements

Inactive Publication Date: 2019-03-19
JINYUBAOLING BIO PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the SVV-specific monoclonal antibody prepared by the Canadian BioVet company has developed a SVV competitive ELISA detection method, which is a relatively mature detection kit, but it is expensive, the purchase cycle is long, and the operation process and result calculation are relatively complicated, which is difficult for operators. It is not suitable for pig farms to detect the infectivity of pigs

Method used

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  • Seneca virus blocking ELISA (enzyme-linked immuno sorbent assay) antibody detection kit
  • Seneca virus blocking ELISA (enzyme-linked immuno sorbent assay) antibody detection kit
  • Seneca virus blocking ELISA (enzyme-linked immuno sorbent assay) antibody detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of Seneca virus blocking ELISA antibody detection kit

[0021] 1. Preparation of Seneca virus antigen

[0022] Select PK-15 cells (purchased from China Veterinary Drug Control Institute) that are full of monolayers, and inoculate Seneca virus (isolated and identified by the seed virus room of the National Engineering Laboratory of Veterinary Vaccines of Jinyu Baoling Biopharmaceutical Co., Ltd.) at a concentration of 2%. Cultivation), when the CPE (cytopathic change) reaches 85%-90%, harvest the venom, freeze and thaw twice at -20°C, add 6% PEG, stir for 4 hours, let stand overnight, and use 8000rpm / min the next day, Centrifuge for 20 minutes, discard the supernatant, and resuspend the pellet; equilibrate the purification column with equilibrium solution (0.02M disodium hydrogen phosphate, 0.15M sodium chloride, pH7.4) for 5-10 column volumes, and select the sample pump to carry out Load the sample, then use the balance solution to elute, and coll...

Embodiment 2

[0037] The usage method of embodiment 2 Seneca virus blocking ELISA antibody detection kit

[0038] 1. Take the kit obtained in Example 1 out of the refrigerator at 4°C, and return to room temperature after about 15 minutes. Dilute the 20-fold wash solution to 1-fold use concentration with purified water.

[0039] 2. Take out the microtiter plate. The unpacked microtiter plate should be stored at 4°C and should be used within 7 days.

[0040] 3. Use the sample diluent to dilute the serum, and dilute the serum to be tested 10 times. The standard positive serum and standard negative serum do not need to be diluted.

[0041] 4. Adding samples: Add standard positive serum to the 1-2 wells of the first row of the microtiter plate, add standard negative serum to the 3rd to 4th wells, and use the 5th to 6th wells as blank controls. Add the serum to be tested to the remaining wells, add 1 well for each serum, do not repeat, shake gently to mix.

[0042] 5. Incubation: Seal the mic...

Embodiment 3

[0052] Example 3 Repeatability Experiment of Seneca Virus Blocking ELISA Antibody Detection Kit

[0053] Select 57 Seneca virus positive sera and 46 negative sera, use the same batch of Seneca virus blocking ELISA antibody detection kit, according to the method of use in Example 2, repeat the detection under the same conditions at different times three times, and the results are shown in Table 1. The results were consistent, indicating that the kits had good reproducibility.

[0054] [Table 1]

[0055]

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PUM

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Abstract

The invention provides a blocking ELISA (enzyme-linked immuno sorbent assay) kit for detecting Seneca virus serum antibodies, a preparation method of the kit and a use method of the kit. The Seneca virus blocking ELISA antibody detection kit is excellent in sensitivity and specificity, the adopted blocking ELISA technology belongs to open operations, technical requirements are less strict, operation steps are simple, convenient and rapid, and the kit can be widely popularized and applied.

Description

technical field [0001] The invention belongs to the field of biological detection, and more specifically, the invention relates to a kit for detecting Seneca virus antibody, a preparation method and a use method of the kit. Background technique [0002] Seneca virus (Seneca valley virus, SVV), also known as Senecavirus A (Senecavirus A, SVA), is the only member of the family Picornaviridae and the genus Senecavirus. SVA, like other picornaviridae members, has a non-encapsulated capsid with a diameter of about 25nm-30nm, which is icosahedral in symmetry. The SVA genome is about 7.2kb in length and is a single-stranded positive-sense RNA with a unique open reading frame (ORF); the ORF is flanked by a 5' untranslated region (about 668 nucleotides) and a 3' untranslated region (about 68 nucleotides), with a poly(A) tail after the 3' untranslated region. The virus was first identified in PER.C6 cell cultures in 2002 and was identified as a contaminant in the cell culture medium...

Claims

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Application Information

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IPC IPC(8): G01N33/535G01N33/569
CPCG01N33/535G01N33/56983
Inventor 杜晓悦任旭荣李超华张妍刘超闫晓焕刘泽
Owner JINYUBAOLING BIO PHARMA CO LTD
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