Anti-vibrio parahaemolyticus chicken yolk antibody, preparation method and application thereof

A technology of chicken egg yolk antibody and hemolytic vibrio, which is applied in the preparation and purification of chicken egg yolk antibody and egg yolk antibody product field. effect and other problems, to achieve the effect of low cost, good specificity and high titer

Active Publication Date: 2009-01-14
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

TDH gene and TRH gene are not single genes, among them, the homology of TDH gene family reaches more than 80%, while the homology of TRH gene and TDH gene is 68%, so a pair of primers cannot detect all TDH and TRH genes
Some scholars found in the study of cases that some pathogenic Vibrio parahaemolyticus urease were positive while KP - , and believed that urease was the main cause of the disease, but related research results showed that most of the urease-positive strains carried TDH and TRH at the same time, so the synergistic effect of TDH and TRH in the pathogenesis of urease-positive Vibrio parahaemolyticus cannot be ruled out
Studies have shown that Vibrio parahaemolyticus in non-clinical samples has only 1% to 2% Kp + parahaemolyticus, but because the toxin-producing conditions of Vibrio parahaemolyticus will be affected by such as the environment of the digestive tract or food matrix, the current health standard does not only use KP as the basis for judgment, but requires that the detection of Vibrio parahaemolyticus should not be detected bacteria

Method used

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  • Anti-vibrio parahaemolyticus chicken yolk antibody, preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0045] Example 1: Preparation of Vibrio parahaemolyticus bacterium antigen

[0046] Inoculate the strain of Vibrio parahaemolyticus into 2% NaCl peptone liquid medium, expand the culture in a shaker at 150r / min at 30°C for 18h, take the culture solution and centrifuge at 4500r / min for 15min to obtain the bacteria, wash the bacteria with sterile physiological saline The suspension of Vibrio parahaemolyticus was obtained by inoculating for 3 times.

Embodiment 2

[0047] Embodiment 2: the inactivation of Vibrio parahaemolyticus bacterium antigen

[0048] Add formaldehyde with a final volume concentration of 0.4% to the bacterial suspension, let stand at 4°C for 12 hours, then centrifuge at 4500 r / min for 15 minutes to remove formaldehyde, and wash the bacteria 3 to 5 times with physiological saline. Adjust the concentration of the bacterial suspension and store it at 4°C for later use.

Embodiment 3

[0049] Example 3: Immunization of hens

[0050] 23-week-old healthy Roman brown layer hens were selected and raised in isolation. Take the spare inactivated bacterial suspension and add an equal volume of Freund's complete adjuvant to make the antigen concentration 5×10 7 cfu / mL and fully emulsified, and then immunized with an immunization dose of 1 mL / bird. The first booster immunization was carried out after an interval of 3 weeks, and the second booster immunization was carried out at an interval of 3 weeks. Freund's incomplete adjuvant was used for the booster immunization. Collect immunized eggs and store them at 4°C.

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Abstract

The invention discloses anti-vibrio parahaemolyticus chicken egg yolk antibody and the preparation method. The anti-vibrio parahaemolyticus chicken egg yolk antibody is prepared and obtained through the steps of preparation and inactivation of vibrio parahaemolyticus antigen, immunization of hens, collection of immunized eggs, coarse extraction of anti-vibrio parahaemolyticus chicken egg yolk antibody from the egg yolk liquid of the immunized eggs and purification. The invention also provides the application of the anti-vibrio parahaemolyticus chicken egg yolk antibody in food safety detection reagents and disease immune diagnostic reagents related with the preparation of vibrio parahaemolyticus and medicines and healthy products or feed additives for preventing and curing the related diseases of vibrio parahaemolyticus. The anti-vibrio parahaemolyticus chicken egg yolk antibody (IgY) prepared and provided by the invention has the advantages that the specificity and the purity are high, the anti-vibrio parahaemolyticus chicken egg yolk antibody has good effects when being used for preparing immunology testing reagents and medicines and feed additives for treating the related diseases of vibrio parahaemolyticus, the cost is low, the production volume is high, and the anti-vibrio parahaemolyticus chicken egg yolk antibody is easy to be industrialized.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a preparation and purification method and application of an egg yolk antibody product with antibody activity—chicken egg yolk antibody against Vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus is a halophilic bacterium widely distributed in oceans and salt lakes. It is one of the most common pathogens of food poisoning in summer. In recent years, foodborne diseases caused by Vibrio parahaemolyticus have always It is the primary cause of primary diseases and seriously threatens people's health. At the same time, vibriosis is also one of the common diseases in mariculture industry. Vibrio parahaemolyticus can infect many kinds of fish and shrimps and cause a large number of deaths, resulting in huge economic losses. In recent years, the intensive culture of seawater fish and shrimp, especially cage culture has become the main culture mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02C07K1/34C07K1/18C07K1/14A61K39/40A61K9/08A61K9/14A61K9/20A61K9/48A61P31/04A23L1/29A23K1/16G01N33/569A23K10/20A23K20/10A23L33/00
Inventor 钟青萍王斌何彬斌蒲俊方祥廖振林
Owner SOUTH CHINA AGRI UNIV
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