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Kit for detecting canine distemper virus blocking ELISA (Enzyme Linked Immunosorbent Assay) antibody of mink, fox and raccoon dog

A technology for canine distemper virus and raccoon dog distemper, applied in raccoon dog distemper virus (CDV) antibody detection kit, fox, ELISA kit for detecting antibody in animal quarantine, fox, raccoon dog distemper virus blocking ELISA, in the field of mink detection, can solve the problems of being unsuitable for a variety of herds, taking a long time, and being unsuitable, achieving reliable results, strong practicability, and good safety

Inactive Publication Date: 2013-12-11
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Serum neutralization test takes a long time, usually 3-4 days, and is not suitable for large-scale serological surveys; while indirect enzyme-linked immunosorbent assay has strict requirements on animal species, and is not suitable for the detection of multiple species of herds

Method used

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  • Kit for detecting canine distemper virus blocking ELISA (Enzyme Linked Immunosorbent Assay) antibody of mink, fox and raccoon dog
  • Kit for detecting canine distemper virus blocking ELISA (Enzyme Linked Immunosorbent Assay) antibody of mink, fox and raccoon dog
  • Kit for detecting canine distemper virus blocking ELISA (Enzyme Linked Immunosorbent Assay) antibody of mink, fox and raccoon dog

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Determination of Optimum Conditions for the Blocking ELISA Kit of Canine Distemper Virus Antibody

[0034] 1. Determination of antigen coating concentration and blocking antibody dilution

[0035] Canine distemper virus was coated with 38.4mg / L, the blocking antibody was diluted 1:100, 1:200, 1:400, 1:800, and indirect ELISA was performed. The results showed that the working concentration of the identified antigen was 9.6mg / L , The dilution of the blocking antibody was 1:400 (see Table 1).

[0036]

[0037] 2. Determination of the dilution of the serum to be tested

[0038]The mink serum was diluted 1:2.5; 1:5; 1:10; 1:20; 1:40; 1:80, and the indirect blocking ELISA was performed according to the optimized conditions. At 10 dilutions, the inhibition rate of immune serum was above 50%, while that of non-immune serum was below 25%. Therefore, the serum dilution was stipulated as 1:10 (see Table 2).

[0039]

Embodiment 2

[0041] Detection of canine distemper virus positive serum titers in 4 mink sera by using canine distemper virus blocking ELISA method

[0042] The main operation method is as follows:

[0043] Canine distemper virus was diluted with carbonate buffer (pH9.6, 0.05 mol / L) and added to a 96-well microtiter plate, 100 μL / well, coated at 37°C for 3 hours, and washed 3 times with a PBST plate washer ; Block with 2% BSA, 200 μL / well, block for 2 hours at 37°C, and wash 3 times with PBST plate washer; dilute mink serum to be tested with PBST and add it to the microtiter plate, 50 μL / well, incubate at 37°C for 30 minutes , add an equal amount of blocking antibody diluted according to the working concentration and incubate for 1 hour, then wash the plate with PBST for 3 times; dilute the goat anti-rabbit enzyme-labeled antibody with PBST, 100 μL / well, incubate at 37°C for 1 hour, and wash the plate with PBST Machine washed 3 times; finally add TMB chromogenic solution, 100 μL / well, an...

Embodiment 3

[0047] Comparison of Canine Distemper Virus Blocking ELISA and Canine Distemper Antibody Neutralization Test

[0048] The main method is as follows:

[0049] Canine distemper virus blocking ELISA method and canine distemper antibody neutralization test were carried out on 30 CDV immune positive sera respectively. The test results showed that the overall coincidence rate of this method was 88.6%. showed good adaptability (see Table 4).

[0050]

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PUM

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Abstract

The invention relates to a kit for detecting a canine distemper virus blocking ELISA (Enzyme Linked Immunosorbent Assay) antibody of a mink, a fox and a raccoon dog. The kit is characterized by comprising an ELISA batten, a blocking antibody, an enzyme-labeled antibody, a substrate color developing solution, a stop solution, positive control, negative control and ELISA envelope antigen, wherein the ELISA envelope antigen adopts the canine distemper virus with the preservation number of CGMCCNO.4858. The kit for detecting the canine distemper virus blocking ELISA antibody which is developed by using an ELISA technology provides a detection method which is accurate, rapid and good in safety, and can be used for screening a large number of canine distemper virus antibodies.

Description

technical field [0001] The present invention relates to a kit for detecting antibodies against canine distemper virus (CDV) in mink, fox and raccoon dogs. Specifically, the present invention particularly relates to an ELISA kit for detecting antibodies in animal quarantine and its application. The invention discloses a blocking ELISA method for detecting canine distemper virus in mink, fox and raccoon dog, belonging to the field of biotechnology. Background technique [0002] Canine distemper virus (CDV) belongs to Paramyxoviridae (Paramyxoviridae), Paramyxovirinae (Paramyxovirinae), Morbillivirus (Morbillivirus). ) is an acute, highly contagious infectious disease that can cause a large number of susceptible animals such as dogs, foxes, raccoon dogs, and minks to become ill, with a mortality rate of 30-80%, and ferrets as high as 100%. The hemagglutinin protein (H protein), which constitutes the main component of the CDV virion envelope, is the main target protein of the b...

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 易立程世鹏
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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