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ELISA (enzyme-linked immuno sorbent assay) detection kit for porcine parvovirus IgM (immunoglobulin m) antibodies as well as preparation method and application of ELISA detection kit

A technology for detecting kits and parvoviruses, applied in measuring devices, instruments, scientific instruments, etc., can solve the problem of rarely detecting PPVIgM antibodies, achieve the effect of improving sensitivity and specificity, overcoming low sensitivity and strong specificity

Inactive Publication Date: 2013-10-23
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods can detect PPV or antibody levels, they rarely detect PPV IgM antibodies

Method used

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  • ELISA (enzyme-linked immuno sorbent assay) detection kit for porcine parvovirus IgM (immunoglobulin m) antibodies as well as preparation method and application of ELISA detection kit
  • ELISA (enzyme-linked immuno sorbent assay) detection kit for porcine parvovirus IgM (immunoglobulin m) antibodies as well as preparation method and application of ELISA detection kit
  • ELISA (enzyme-linked immuno sorbent assay) detection kit for porcine parvovirus IgM (immunoglobulin m) antibodies as well as preparation method and application of ELISA detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Preparation of ELISA kit for detection of porcine parvovirus IgM antibody

[0053] 1. The porcine IgM monoclonal antibody of this embodiment was purchased from MP Biomedicals, Incorporated;

[0054] 2. the preparation of horseradish peroxidase (HRP)-anti-PPV antibody enzyme conjugate, comprises the steps:

[0055] 1) Routinely immunize rabbits with the purified PPV antigen, and when the ELISA titer of the rabbit serum reaches 1:800 or more, take the serum;

[0056] 2) Precipitating and purifying the serum prepared in step 1) through ammonium sulfate to obtain purified serum;

[0057] 3) HRP-labeling the purified serum prepared in step 2) by an improved sodium periodate oxidation method to obtain a horseradish peroxidase (HRP)-anti-PPV antibody enzyme conjugate;

[0058] 4) Add 0.1-10% bovine serum albumin, 0.1-10% casein and 50% neutral glycerol to the enzyme marker (i.e. horseradish peroxidase (HRP)-anti-PPV antibody enzyme conjugate), After measuring th...

Embodiment 2

[0063] Example 2 Detection method of porcine parvovirus ELISA antibody detection kit

[0064] Utilize the detection method of porcine parvovirus IgM antibody ELISA detection kit of the present invention, comprise the steps:

[0065] 1) Dilute the sample to be tested 100 times with the sample diluent, add it to the wells of the microplate plate evenly coated with porcine IgM monoclonal antibody, add 100 μl to each well, and set a positive control group, a negative control group and a blank control group at the same time 100 μl positive control solution was added to the positive control group, 100 μl negative control solution was added to the negative control group, 100 μl sample diluent was added to the blank control group, and 3 wells were added in parallel for each sample and control;

[0066] 2) After adding the samples, incubate the ELISA plate at 37° C. for 1 hour, wash the plate with washing solution 3 times, and spin dry. The washing solution is prepared by diluting 10...

Embodiment 3

[0072] Example 3 Specificity and sensitivity test of the kit of the present invention

[0073] 1. Specific detection

[0074] According to the detection method described in Example 2, the kits prepared in Example 1 were used to detect respectively the early serum of porcine PPV vaccine immunity, the early serum of porcine PPV infection, the purified porcine PPV IgG antibody, and the early serum of porcine circovirus type 2 infection , porcine parvovirus antibody positive serum, porcine pseudorabies virus positive serum, swine fever virus antibody positive serum and porcine PPV negative serum.

[0075] Dilute the sample to be tested 100 times with the sample diluent, add it to the wells of the microplate plate evenly coated with 0.1μg-1μg / well of porcine IgM monoclonal antibody, add 100μl to each well, and set up a positive control group and a negative control group in parallel 100 μl of positive control solution was added to the positive control group, 100 μl of negative co...

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Abstract

The invention relates to an ELISA (enzyme-linked immuno sorbent assay) detection kit for porcine parvovirus IgM (immunoglobulin m) antibodies as well as a preparation method and application of the ELISA detection kit. The ELISA detection kit for the porcine parvovirus IgM antibodies comprises an ELISA plate coating a porcine IgM monoclonal antibody, sealing fluid, a sample diluent, a detection antigen, an enzyme conjugate, a concentrated cleaning solution, a zymolyte solution A, a zymolyte solution B and a stop solution, wherein the detection antigen is a purified porcine parvovirus, and the enzyme conjugate is a horse-radish peroxide enzyme-anti-PPV (porcine parvovirus) antibody enzyme conjugate. The specificity of the ELISA detection kit provided by the invention reaches 100% and the sensitivity of the ELISA detection kit reaches up to 1:800, so that the ELISA detection kit can be used for performing early diagnosis on PPV infection of a swinery.

Description

technical field [0001] The invention relates to a detection kit, in particular to a porcine parvovirus (PPV) IgM antibody ELISA detection kit, a preparation method and an application thereof. Background technique [0002] Porcine parvovirus infection (PPI), also known as porcine reproductive disorder, is a reproductive disorder of pigs caused by porcine parvovirus (PPV), and it is also one of the main diseases that cause reproductive disorder in sows. Pregnant sows mainly show abortion, stillbirth, mummified fetuses, weak piglets, etc., and the first-born sows are the most serious, causing huge economic losses to the global pig industry. [0003] Currently, there is no effective drug treatment for PPV infection. In recent years, it has been recognized that the use of vaccines is the only way to prevent PPV infection and increase the reproductive rate of sows. Although inactivated vaccines are the safest, experiments have shown that PPV can be sufficiently attenuated that e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 廖园园漆世华李建秦伟张萍谢红玲温文生
Owner WUHAN CHOPPER BIOLOGY
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