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Test strip card for quantitative detection of classical swine fever virus antibody with double antigen sandwich and double detection line

A double-antigen sandwich, swine fever virus technology, applied in measurement devices, biological tests, material inspection products, etc., can solve the problems of insignificant difference in detection color and low sensitivity, and achieve improved sensitivity, sufficient response, and increased response time. Effect

Active Publication Date: 2021-08-27
武汉观锐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to overcome the shortcomings of the existing colloidal gold rapid detection method for qualitative detection or semi-quantitative and quantitative detection methods with low sensitivity and insignificant detection color difference, and to provide a kind of swine fever with double antigens and double detection lines Virus Antibody Quantitative Detection Test Strip Card and Application

Method used

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  • Test strip card for quantitative detection of classical swine fever virus antibody with double antigen sandwich and double detection line
  • Test strip card for quantitative detection of classical swine fever virus antibody with double antigen sandwich and double detection line
  • Test strip card for quantitative detection of classical swine fever virus antibody with double antigen sandwich and double detection line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The preparation of embodiment 1 swine fever virus E2 and E2a recombinant antigen

[0047] This example provides a preparation method of recombinant antigens of classical swine fever virus E2 and E2a as follows: use IPTG to induce recombinant strains producing E2 and E2a proteins respectively, and the expression products are purified to obtain recombinant antigens of classical swine fever virus E2 and classical swine fever virus E2a respectively recombinant antigen.

[0048] (1) The preparation method of CSFV E2 recombinant antigen is as follows:

[0049] The classical swine fever virus E2 antigen protein genome sequence was searched in the NCBI database, and GenBank: FJ598611.1 (Classical swine fever virus strain C-strain-ZJ E2glycoprotein gene) was selected as the target gene. The specific gene sequence is shown in SEQ ID NO: 1.

[0050] Use the Primer premier5 software to design the PCR primers for amplifying the E2 gene, and the sequences of the primers are as follo...

Embodiment 2

[0068] The assembly of embodiment 2 swine fever virus antibody quantitative detection test paper card

[0069] The embodiment of the present invention provides a preparation method of a test paper card for quantitative detection of classical swine fever virus antibody based on double detection line technology, the method comprising: sample pad, classical swine fever virus E2 protein gold standard pad, rabbit IgG gold standard pad, water-absorbing pad , the nitrocellulose membrane coated with the double detection line of the classical swine fever virus E2a protein and the quality control line of goat anti-rabbit IgG, and the PVC rubber plate according to the image 3 Assembled sequentially to obtain the test paper card for quantitative detection of classical swine fever virus antibody.

[0070] Wherein, refer to Example 1 for the preparation method of CSFV E2 and E2a recombinant antigens.

[0071] Preparation method of classical swine fever virus E2 protein gold label pad: tak...

Embodiment 3

[0079] The preparation of embodiment 3 swine fever virus antibody quantitative detection test paper card

[0080] The embodiment of the present invention provides a preparation method of a test paper card for quantitative detection of classical swine fever virus antibody based on double detection line technology, the method comprising: sample pad, classical swine fever virus E2 protein gold standard pad, rabbit IgG gold standard pad, water-absorbing pad , the nitrocellulose membrane coated with the double detection line of the classical swine fever virus E2a protein and the quality control line of goat anti-rabbit IgG, and the PVC rubber plate according to the image 3 Assembled sequentially to obtain the test paper card for quantitative detection of classical swine fever virus antibody.

[0081] Wherein, the preparation method of the recombinant antigen of classical swine fever virus E2 and E2a refers to Example 2.

[0082] Preparation method of classical swine fever virus E...

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Abstract

The invention belongs to the field of animal immune detection, and in particular relates to a test paper card for quantitative detection of swine fever virus antibody with double antigen sandwiches and double detection lines. The test card includes sample pad, classical swine fever virus E2 protein gold standard pad, rabbit IgG gold standard pad, absorbent pad, double detection line coated with classical swine fever virus E2a protein, and nitrocellulose membrane of goat anti-rabbit IgG quality control line and PVC rubber sheet; colloidal gold adopts double-antigen sandwich and double detection line; the classical swine fever virus gold standard pad is coated with the protein antigen secreted by Escherichia coli BL21 / pE2 with the preservation number CCTCC NO: M2018060. E2 antigen and gold-labeled rabbit IgG; the nitrocellulose membrane is coated with the CSFV E2a antigen constructed from the protein antigen secreted by Escherichia coli BL21 / pE2a with the preservation number CCTCC NO: M2018059, constituting a double detection line area and The quality control line region is composed of goat anti-rabbit IgG.

Description

technical field [0001] The invention belongs to the technical field of animal immunity application. In particular, it relates to a test paper card for quantitative detection of swine fever virus antibody with a double antigen sandwich and a double detection line. [0002] The test paper card of the invention can be used for real-time and rapid quantitative detection of the antibody to the classical swine fever virus, and is of great significance to the monitoring of the health level of pig herds and the antibody level after immunization. Background technique [0003] Classical Swine fever (CSF) is an acute, febrile, high-contact porcine infectious disease caused by classical swine fever virus (CSFV). Swine fever virus is mainly transmitted through the digestive tract and respiratory tract. It spreads quickly, has high morbidity and mortality, and brings huge economic losses to the global pig industry. It is listed as 16 types of A by the World Organization for Animal Health...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56983G01N33/6854G01N33/6893G01N2333/183
Inventor 刘锡玲龙兴权代娟李佳黄双龙李筱雯况世昌吴贝
Owner 武汉观锐生物科技有限公司
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