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Quantitative detection method and kit of specific IgA anti-body for porcine epidemic diarrhea virus in sow colostrum

A technology for porcine epidemic diarrhea and breeding sows, applied in the biological field, can solve the problems of not being able to quantitatively detect IgA antibodies, not being suitable for on-site detection, affecting childbirth or lactation, etc., to achieve optimal immunization schemes, short time consumption, and predictive protection Effect

Inactive Publication Date: 2019-08-06
北京测易生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] CN104330558A discloses an ELISA method for detecting IgA antibodies in sow blood, which can qualitatively detect IgA antibodies in sow blood by indirect ELISA method, and has good specificity, but cannot quantitatively detect IgA antibodies in sow blood level, and since it involves postpartum or prepartum sow blood collection, it may cause some stress to the sow, and even affect farrowing or lactation
In addition, the ELISA method requires professional instruments (microplate readers, water baths, etc.), and requires experimental personnel with certain professional qualities to carry out in relatively professional laboratories, but it is not suitable for on-site testing.

Method used

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  • Quantitative detection method and kit of specific IgA anti-body for porcine epidemic diarrhea virus in sow colostrum
  • Quantitative detection method and kit of specific IgA anti-body for porcine epidemic diarrhea virus in sow colostrum
  • Quantitative detection method and kit of specific IgA anti-body for porcine epidemic diarrhea virus in sow colostrum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of colloidal gold test strip and test card for detecting porcine epidemic diarrhea virus-specific IgA antibody level in sow colostrum

[0043] The colloidal gold test strip or test card provided in this example can be used for rapid and on-site detection of porcine epidemic diarrhea virus-specific IgA antibody levels in colostrum of sows, and can be used to detect vaccine quality and quality control during vaccine production, etc. .

[0044] 1.1. Preparation of colloidal gold test strips and test cards

[0045] like figure 1 Shown is a schematic diagram of the structure of the colloidal gold test strip, wherein figure 1 Panel A in the middle is a schematic diagram of the front structure of the colloidal gold test strip. figure 1 Panel B in the middle is a schematic diagram of the side structure of the colloidal gold test strip.figure 1 Panel C in the middle is the chromatographic direction of the colloidal gold test strip, according to figure ...

Embodiment 2

[0060] Example 2: Kit for rapid quantitative detection of porcine epidemic diarrhea virus-specific IgA antibody in sow colostrum

[0061] The kit in this example is based on the colloidal gold immunochromatography technology, and the supporting KRD-105 immunochromatography analyzer (model KRD-105, purchased from Beijing Kuairuidu Technology Co., Ltd.) can realize rapid detection of sow colostrum. Levels of epidemic diarrhea virus-specific IgA antibodies.

[0062] 2.1. The kit includes:

[0063] 1) Colloidal gold test cards: several (20 in this embodiment), prepared according to the method described in Example 1, are individually packaged in aluminum foil bags. Each aluminum foil bag contains a test card and a pack of desiccant;

[0064] 2) diluent: several bottles (20 bottles in this embodiment), each bottle contains 630 μl of diluent, wherein the diluent is 10mM PBS (formula: NaH 2 PO 4 2H 2 O 0.296g, Na 2 HPO 4 12H 2 (2.9g, dissolved in double distilled water to 1000...

Embodiment 3

[0080] Example 3: Detection of porcine epidemic diarrhea virus-specific IgA antibody level in sow colostrum

[0081] This embodiment utilizes the kit of embodiment 2 to detect, comprising the following steps:

[0082] 1), the preparation of sow colostrum sample to be tested: collect the colostrum of postpartum sow within 48 hours, make a good mark; Because sow colostrum is relatively viscous, need to use diluent (same as in Example 2) before detection The diluent) is diluted according to a ratio of 1:10 to prepare a sample of sow colostrum to be tested; for example, 70 μl of colostrum is added to 630 μl of diluent to prepare a sample of sow colostrum to be tested diluted at a ratio of 1:10 ;

[0083] 2), take out a sufficient number of test cards from the test kit in Example 2, open the package, and number them according to the order of the samples; add 70 μl of the diluted sow colostrum sample to be tested to the sample hole of the test card successively, and time 20 minute...

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Abstract

The invention discloses a quantitative detection method and kit of specific IgA anti-body for porcine epidemic diarrhea virus in sow colostrum, and belongs to the technical field of biology. The method and kit based on a colloidal gold immunochromatography technique are simple and rapid in operation and especially suitable for onsite detection. The colostrum of a sow after birth is collected without wound for detection, the sow is stimulated less and avoided from influence on birth or lactation, and the titer level of the specific IgA anti-body for porcine epidemic diarrhea virus in sow colostrum is detected quantitatively, protective effectiveness of sow colostrum for piglings can be further distinguished, the immunization scheme for anti porcine epidemic diarrhea virus of the sow can beoptimized further, and high-quality vaccines can be obtained by screening.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a quantitative detection method and kit for porcine epidemic diarrhea virus-specific IgA antibody in sow colostrum. Background technique [0002] Porcine epidemic diarrhea is an acute intestinal infectious disease that seriously threatens the health of newborn piglets. It is caused by porcine epidemic diarrhea virus (Porcine Epidemic Diarrhea Virus, PEDV). It is characterized by vomiting, diarrhea, and dehydration. Clinical changes and symptoms are very similar to porcine transmissible gastrointestinal disease. It first appeared in the UK in 1971, and the disease occurred successively in my country in the early 1980s. The fatality rate of piglets often exceeds 50%, which is a major disease affecting the breeding industry. [0003] The prevention of porcine epidemic diarrhea mainly adopts vaccines at present, and vaccines are immunized to pregnant sows. Pigs generally produce IgA ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983
Inventor 郑金来吴园园郝金宝卢小雨张卉李卫丽周博李丽燕
Owner 北京测易生物科技有限公司
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