Muscovy duck gosling plague latex particle agglutination reagent and preparation method thereof

A technology for latex agglutination and goose blast, applied in biochemical equipment and methods, microbe-based methods, antiviral immunoglobulin, etc., can solve the problems of lack of rapid diagnostic reagents and difficulties in clinical diagnosis of muscovy duck and goose blast, and achieve High repeatability and accuracy, long shelf life, and good sensitivity

Active Publication Date: 2012-02-08
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, there is still a lack of commercialized rapid diagnostic reagents for muscovy d

Method used

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  • Muscovy duck gosling plague latex particle agglutination reagent and preparation method thereof
  • Muscovy duck gosling plague latex particle agglutination reagent and preparation method thereof
  • Muscovy duck gosling plague latex particle agglutination reagent and preparation method thereof

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Experimental program
Comparison scheme
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Example Embodiment

[0008] The preparation method of latex agglutination reagent of muscovy duck and gosling plague is as follows:

[0009] Dilute 10% polystyrene latex to 1% with 0.1M pH8.2 glycine buffer (GBS), centrifuge at 6000r / min for 5min, discard the supernatant, resuspend the precipitate to the original volume with 0.1M pH8.2GBS, add Equal volume of anti-GPV monoclonal antibody (protein concentration 0.5 ~ 1mg / ml), mix thoroughly, put in a 37°C water bath for 40min (shake for a few seconds every 10min), then use 1% bovine serum albumin (BSA) Wash twice with 0.1M pH8.2GBS, centrifuge at 6000r / min for 5min each time, resuspend the pellet in 0.1%BSA-GBS to make the final concentration of latex 1%, and add sodium azide to the final concentration of 0.025%. Latex agglutination reagent for muscovy duck and gosling plague.

[0010] in:

[0011] 1. Preparation of monoclonal antibody against goose parvovirus from Muscovy duck:

[0012] Take out the anti-GPV hybridoma cell line D11 from the liq...

Example Embodiment

[0042] Experimental example

[0043] According to the operation method of latex agglutination test (LPA), the latex agglutination reagent was taken to react with different antigens and normal cell culture fluid respectively. As a result, the latex agglutination reagent only had a specific agglutination reaction with GPV, and did not react with other viruses; GPV embryo fluid, sick duck The highest agglutination titer (LPA titer) of liver and spleen tissue homogenate was 2 6 and 2 2 (table 3).

[0044] Table 3. Identification of GPV antigens using latex agglutination assay (LPA)

[0045]

[0046] Note: GPN is GPV embryo sap poison, GPT is GPV-infected duck liver and spleen homogenate, MPV is Muscovy duck parvovirus, MDRV is Muscovy duck reovirus, ARV is avian reovirus, PMV is duck paramyxovirus, DHV For duck hepatitis virus. The data in the table is LPA titer, "-" means negative.

[0047] 6.2 Rapid diagnosis of gosling plague:

[0048] Experimental example

[0049] Ac...

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Abstract

The invention discloses a muscovy duck gosling plague latex particle agglutination reagent. The muscovy duck gosling plague latex particle agglutination reagent is characterized in preparation steps that: a GPV-resisting hybridoma cell strain D11 is used for preparing a GPV monoclonal antibody, and the GPV monoclonal antibody is used for preparing the muscovy duck gosling plague latex particle agglutination reagent; the GPV-resisting hybridoma cell strain D11 is preserved in CCTCC with a preservation date of April 15th, 2011 and a preservation number of C201120. The invention also discloses applications of latex particle agglutination (LPA) tests and latex particle agglutination inhibition (LPAI) tests established by using the muscovy duck gosling plague latex particle agglutination reagent in the determinations of goose parvovirus antigens, in the diagnosis of muscovy duck gosling plague, in serological investigation, and in immune antibody monitoring.

Description

technical field [0001] The invention relates to a latex agglutination reagent for muscovy duck and goose blast. Background technique [0002] Muscovy duck goose plague is a viral infectious disease caused by goose parvovirus (Goose parvovirus, GPV) in young muscovy ducks less than one month old. , At present, there are occurrences in all Muscovy duck breeding areas across the country. Clinically, the young muscovy ducks are characterized by diarrhea and intestinal mucosa shedding to form embolism, with a morbidity rate of 50% to 70% and a mortality rate of 40% to 65%, causing certain economic losses to the duck industry. Clinically, the disease is not only often co-infected with Muscovy duck parvovirus disease (three-week disease), but also often misdiagnosed as Muscovy duck parvovirus disease by grassroots veterinarians. Since the hyperimmune serum against gosling plague has a good therapeutic effect on the disease, rapid diagnosis is the premise of effectively controllin...

Claims

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Application Information

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IPC IPC(8): G01N33/577C07K16/08C12N5/20C12R1/91
Inventor 陈少莺朱小丽林锋强程晓霞陈仕龙王劭
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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