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

A technology for porcine Japanese encephalitis and parvovirus, which is used in measuring devices, instruments, scientific instruments, etc., can solve the problems of inconvenient operation and inconvenience for non-professional grassroots personnel, and achieves simple operation, high labeling rate, and detection specificity. strong effect

Inactive Publication Date: 2012-07-04
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 washes are required to obtain the result; its operation is still not simple enough, and it is still not convenient for non-professional grassroots personnel to use

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

Examples

Experimental program
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Effect test

preparation example Construction

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

[0027] Inject negative healthy sheep or rabbits 3 to 4 times subcutaneously and intramuscularly with 50μg~100μg SPA / kg body weight, collect blood venously 20 days after the last immunization, measure the serum antibody titer above 1:2000 by ELISA, collect blood from the heart or carotid artery Bloodletting and collection of hyperimmune serum. Take 1 part of serum and 2 parts of PBS solution (pH7.2) and mix well, add an equal volume of saturated ammonium sulfate solution and mix well, put it in a refrigerator at 4°C for 2 hours, centrifuge at 4°C and 10000r / min for 15min, discard the supernatant; Appropriate amount of PBS solution (pH7.2) to dissolve the precipitate, add saturated ammonium sulfate solution to the final concentration of 33%, put it in the refrigerator at 4°C for 2h, centrifuge at 4°C and 10000r / min for 15min, discard the supernatant, and use a small amount of PBS solution (pH7.2) dissolves the precipitate,...

Embodiment 1

[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, which is made of glass wool, 3 is the fiber layer of adsorbed gold standard substance, which is made of glass with colloidal gold labeled SPA Made of cotton, prepare its gold standard SPA glass wool according to the preparation method described in the above specific embodiment 4, 4 is a cellulose membrane layer, made of nitrocellulose membrane, and 5 is a water-absorbing material layer, made of water-absorbing filter paper ; Paste each layer of numbers 2, 3, 4, and 5 on the hard plastic sheet strip 1 from the left end test end to the right, and the fibers at the junction between each other interpenetrate each other; on the nitrocellulose membrane layer 4, 6 uses purification The...

Embodiment 2

[0044] Embodiment two: one step detects the test strip of porcine parvovirus, porcine encephalitis virus antibody, which is basically the same as embodiment one, except that:

[0045] Adsorption gold standard material fiber layer 3 is made with the glass wool of PPV and JEV two kinds of purification virus antigen liquids that are adsorbed with colloidal gold mark, prepares its gold standard purification virus antigen (Xi ) glass wool; on the nitrocellulose membrane layer 4, 7 is the 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 test sample preparation, operation method and result judgment are the same as the operation method in 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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Patent Type & Authority Patents(China)
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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