Test paper strip for detecting Porcine Parvovirus and porcine encephalitis B virus antibody

A technology for porcine Japanese encephalitis and parvovirus, which is applied to measuring devices, instruments, scientific instruments, etc., can solve the problems of inconvenience for non-professional grass-roots personnel, and the operation is not easy enough, and achieves easy operation, high labeling rate, and sensitivity. high effect

Inactive Publication Date: 2009-01-14
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the currently reported dot immunogold diafiltration method can only detect one kind of swine disease antibody at a time, and at least two reagents and two

Method used

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  • Test paper strip for detecting Porcine Parvovirus and porcine encephalitis B virus antibody
  • Test paper strip for detecting Porcine Parvovirus and porcine encephalitis B virus antibody

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0026] 1. Preparation of goat anti- or rabbit anti-SPA IgG:

[0027] 50μg-100μg SPA / kg body weight was injected subcutaneously and intramuscularly into negative healthy sheep or rabbits 3 to 4 times. Blood was collected 20 days after the last immunization, and the serum antibody titer was determined to be above 1:2000 by ELISA. Blood was collected from the heart or carotid artery. Bleeding and collecting its high immune serum Take 1 part of serum and 2 parts of PBS solution (pH 7.2) and mix well, add an equal volume of saturated ammonium sulfate solution and mix well, place in a refrigerator at 4°C for 2h, centrifuge at 4°C, 10000r / min for 15min, and discard the supernatant; Dissolve the precipitate in PBS solution (pH 7.2), add saturated ammonium sulfate solution to its final concentration of 33%, place in a refrigerator at 4°C for 2 hours, centrifuge at 4°C and 10000r / min for 15 minutes, discard the supernatant, and use a small amount of PBS Solution (pH7.2) to dissolve the prec...

Example Embodiment

[0041] Example 1: Test strips for one-step detection of porcine parvovirus and porcine Japanese encephalitis virus antibodies

[0042] See figure 1 with figure 2 In the figure, 1 is the support layer, which is made of hard plastic thin strips, 2 is the sample adsorption fiber layer of the test end, made of glass wool, and 3 is the gold label fiber layer, and the glass is adsorbed with colloidal gold to mark SPA. Made of cotton, the gold-labeled SPA glass wool was prepared according to the preparation method described in the above-mentioned specific embodiment 4. 4 is a cellulose membrane layer, made of nitrocellulose membrane, 5 is a water-absorbing material layer, made of water-absorbing filter paper ; Paste each layer numbered 2, 3, 4, 5 on the hard plastic sheet 1 from the left end to the right test end, and the fibers at the junction between each other cross and penetrate each other; on the nitrocellulose membrane layer 4, 6 is used for purification The detection blots P and ...

Example Embodiment

[0044] Example 2: The test strip for one-step detection of porcine parvovirus and porcine Japanese encephalitis virus antibodies is basically the same as Example 1, except for:

[0045] The gold-labeled fiber layer 3 is made of glass wool adsorbed with colloidal gold-labeled PPV and JEV two purified virus antigen solutions, and the gold-labeled purified virus antigen (Xi) is prepared according to the preparation method described in the above-mentioned specific embodiment 4. ) Glass wool; on the nitrocellulose membrane layer 4, 7 is a control blot C printed with sheep (or rabbit) anti-PPV and JEV IgG solutions, and the two blots are arranged in parallel and combined as "||| ". Others including the preparation of the test sample, the operation method and the result determination, etc. are the same as the operation method in the specific embodiment 6, and will not be repeated.

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Abstract

The invention discloses a test strip that is used for detecting antibodies of porcine parvovirus and porcine encephalitis B virus in one step, which comprises a supporting layer, an adsorptive layer and a protecting layer, and the adsorptive layer consists of a sample fiber layer, an adsorptive gold mark fiber layer, a fibrin membranous layer and a water absorptive material layer sequentially from a testing end; the adsorptive gold mark fiber layer adsorbs any one or two from antigen liquids of gold colloid marked SPA or gold colloid marked pure PPV and JEV; a detecting print and a contrast print are arranged on the fibrin membranous layer, the detecting print is printed with any one or two from the purified PPV and JEV antigen liquids, and the contrast print is printed with an IgG solution of ovine or rabbit anti-SPA or an IgG solution of any one or two viruses from ovine or rabbit anti-PPV and anti-JEV. The test strip of the invention has strong detecting specificity and high sensitivity, avoids additional equipment and reagents, can be operated by every person, and is particularly applicable to the quick specific antibody detection of porcine diseases on the spot.

Description

1. Technical field: [0001] The invention relates to a device for detecting antibodies to porcine reproductive disorder virus infectious diseases, in particular to a test strip for one-step detection of antibodies to porcine parvovirus and porcine encephalitis virus. 2. Technical background: [0002] my country is a big producer and consumer of live pigs. With the number of large, medium and small-scale pig farms and breeding pig farms increasing year by year, pig reproductive disorders have become one of the most important diseases in large-scale pig farms in my country, with an average incidence of more than 30%, causing huge economic losses. losses, and seriously restrict the development of the pig industry. Porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), porcine encephalitis virus (JEV), swine fever virus and pseudorabies virus are important viruses that cause reproductive disorders in pigs. PRRSV causes porcine reproductive and res...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/545G01N33/544
Inventor 张改平肖治军邓瑞广李学伍杨艳艳杨继飞赵东王爱萍邢广旭柴书军刘庆堂
Owner HENAN ACAD OF AGRI SCI
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