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Test paper card for detecting pseudorabies virus gE protein antibody in porcine serum as well as preparation method and application thereof

A technique for pseudorabies virus and porcine pseudorabies, which is applied to a test paper card for detecting pseudorabies virus gE protein antibody in swine serum and the fields of preparation and application, which can solve the problems of high cost and long time, and achieve high sensitivity and detection time. short, simple effects

Inactive Publication Date: 2015-05-06
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, enzyme-linked immunosorbent assay (ELISA) is generally used to detect porcine pseudorabies virus gE protein antibody in China, but due to the particularity of this detection method, it must be completed in a laboratory, which takes a long time and costs high

Method used

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  • Test paper card for detecting pseudorabies virus gE protein antibody in porcine serum as well as preparation method and application thereof
  • Test paper card for detecting pseudorabies virus gE protein antibody in porcine serum as well as preparation method and application thereof

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Experimental program
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Effect test

Embodiment 1

[0039] Such as figure 1 and figure 2 As shown, a test paper card for detecting pseudorabies virus gE protein antibody in pig serum, the test paper card is composed of an immune colloidal gold test strip and a card body. The card body includes a support backplane (7), a detection window hole (4) and a sample injection hole (2); the immunocolloidal gold test strip includes a sample pad (6), a gold standard binding pad (8), a nitrocellulose membrane ( 9), absorbent pad (10), bottom plate (11), nitrocellulose membrane (9) is provided with a test line (3) coated with porcine pseudorabies virus strain E A and a quality control line coated with goat anti-mouse IgG (5). The connection relationship is: the nitrocellulose membrane (9) is pasted on the bottom plate (11), and the gold label binding pad (8) and the sample pad (6) are pasted on one end of the nitrocellulose membrane (9), and the absorbent pad (10) Paste on the other end of the nitrocellulose membrane 9; the joints (1-3 ...

Embodiment 2

[0045] The preparation of the test card is based on the detection technology of colloidal gold immunochromatography. In this study, the production of the antibody test card for the detection of pseudorabies virus gE protein in pig serum is a competitive method.

[0046] A preparation method of a test paper card for detecting pseudorabies virus gE protein antibody in pig serum, the steps of which are:

[0047] A. Preparation and purification of porcine pseudorabies virus: take out pseudorabies virus E A strain stored at -78~-82°C, infect a single layer of pK-15 cells, extract virus DNA, and inoculate at 0.5 MOI after PCR identification For pK-15 cells, 90% of the cytopathic cells appear to collect the virus. The virus liquid is repeatedly frozen and thawed 2 to 4 times, inactivated at 56°C, the virus liquid is centrifuged at 8000rpm, the precipitate is discarded, and the centrifuged supernatant is put into a clean In a beaker, under the stirring of a magnetic stirrer, add ammon...

Embodiment 3

[0060] A kind of test paper card of pseudorabies virus gE protein antibody in pig serum is used in detecting the application of pseudorabies virus gE protein antibody in pig serum, and its steps are:

[0061] A. Sample processing: Blood collection from the anterior vena cava of pigs, place the blood sample at 4°C for 1 hour, use a pipette gun to take out the precipitated serum and put it into a clean EP tube, numbered;

[0062] B. Detection: 1) Take out the sealed bag from the 4°C refrigerator, take out the test card from the sealed bag, and place it flat at room temperature (20°C~25°C) to return to normal temperature. 2) Dilute the serum to be tested twice with the sample diluent: mix 60 μl of the sample diluent and 60 μl of the serum to be tested evenly; 3) Take out 100 μl of the above mixture and slowly drop it into the sample well. Place the strips flat on the table and observe the results within 30 minutes.

[0063] C. Result judgment: when the test sample is...

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Abstract

The invention discloses a test paper card for detecting a pseudorabies virus gE protein antibody in porcine serum as well as a preparation method and application thereof. The test paper card is formed by an immuno-colloidal gold test strip and a card body; a nitrocellulose membrane is adhered to a bottom plate of the immuno-colloidal gold test strip; a gold-marking binding pad and a sample pad are adhered to one end of the nitrocellulose membrane, and a water absorbing pad is adhered to the other end of the nitrocellulose membrane; the nitrocellulose membrane is provided with a porcine pseudorabies virus EA strain coated detection line and a goat anti-mouse IgG coated quality control line, wherein the detection line and the quality control line are parallel to each other, the detection line is arranged near to one end of the gold-marking binding pad, and the quality control line is arranged near to one end of the water absorbing pad; the immuno-colloidal gold test strip is arranged in the test paper card body; and the detection line and the quality control line are arranged in positions which correspond to detection holes of the test paper card. The test paper card disclosed by the invention is used for detecting the pseudorabies virus gE protein antibody in the porcine serum rapidly and has the advantages of simple structure, convenience of operation, high sensitivity and clear result.

Description

technical field [0001] The invention relates to the technical field of animal virology and immunology detection, more specifically to an immune colloidal gold test paper card for detecting pseudorabies virus gE protein antibody in pig serum, and also relates to a method for detecting pseudorabies virus gE protein antibody in pig serum. The preparation method of the immune colloidal gold test paper card for rabies virus gE protein antibody also relates to an application of the immune colloidal gold test paper card for detecting pseudorabies virus gE protein antibody in pig serum. Background technique [0002] Porcine pseudorabies (PR) is an acute infectious disease caused by pseudorabies virus (PRV). Infected pigs are clinically characterized by elevated body temperature and neurological symptoms in newborn piglets. Adult pigs are often recessively infected, and pregnant sows can cause abortion, stillbirth and respiratory symptoms after infection. The disease is widely dist...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/56983G01N33/577
Inventor 吴斌李慧但汉并彭忠胡睿铭汤细彪董晓辉何启盖陈焕春
Owner HUAZHONG AGRI UNIV
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