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Detection kit, preparation method and application of hemagglutination-inhibiting antibody of Aviobacterium gallinarum

A technique for inhibiting hemagglutination of Avium nasotrachea and hemagglutination, which is applied in the field of detection kits, preparation and application of hemagglutination inhibitory antibodies of Avium nasitrachea, and can solve the problems of inability to detect ORT antibodies, etc.

Active Publication Date: 2015-08-12
YUNNAN ANIMAL SCI & VETERINARY INST
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Problems solved by technology

[0005] The purpose of the present invention is to overcome the deficiency that the above-mentioned existing imported ORT indirect ELISA antibody detection kit cannot detect the ORT antibody in chicken egg yolk, and to invent a detection kit for the hemagglutination inhibition antibody of Aviobacterium nasotrachea

Method used

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  • Detection kit, preparation method and application of hemagglutination-inhibiting antibody of Aviobacterium gallinarum
  • Detection kit, preparation method and application of hemagglutination-inhibiting antibody of Aviobacterium gallinarum
  • Detection kit, preparation method and application of hemagglutination-inhibiting antibody of Aviobacterium gallinarum

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Embodiment

[0036] (1) Preparation of ORT hemagglutinating antigen: Ornithobacterium rhinotracheale YN12091 (strain preservation number: CCTCC No.: M2013318, referred to as ORT) Yunnan isolate YN12091 (strain preservation number: CCTCC No.: M2013318) , the strains were inoculated on nutrient agar medium, placed in 5% CO 2Incubate in an incubator at 37°C for 24 hours, wash the colonies on the nutrient agar medium with PBS (0.1M, Ph6.3), centrifuge at 4000rpm for 10min, discard the supernatant, resuspend the bacteria sludge with PBS, and centrifuge again. Repeat the centrifugation and resuspension steps to wash the ORT bacteria 3 times, discard the supernatant after the third centrifugation, resuspend the bacteria sludge with PBS, and adjust its concentration to 15mg / mL (the protein concentration is measured by an ultra-micro ultraviolet spectrophotometer), Add thimerosal at a final concentration of 0.01% (m / v), and store at 4°C for later use.

[0037] (2) Preparation of aldehydated sheep ...

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Abstract

The invention discloses a detection kit for detecting ornithobacterium rhinotracheale hemagglutinin inhibitor antibody, a preparation method, and applications thereof, and especially relates to a detection kit for detecting hemagglutinin inhibitor antibody of the ornithobacterium rhinotracheale YN12091 whose bacterial strain preservation number is CCTCC No: M2013318, being named ORT for short, a preparation method, and applications thereof. The preparation method comprises following steps: choosing ORT high hemagglutinin activity bacterial strains, culturing and washing to prepare ORT hemagglutinin antigens; collecting sheep blood, preparing sheep erythrocytes from the collected sheep blood through a washing and centrifugation method, fixing the sheep erythrocytes with glutaraldehyde and then tanning the sheep erythrocytes with tannic acid so as to prepare hydroformylated sheep erythrocytes; immunizing chickens for three times with an ORT bacterial suspension, then preparing ORT standard positive serum through centrifugation of the collected blood; assembling the prepared ORT hemagglutinin antigens, hydroformylated sheep erythrocytes, and ORT standard positive serum to establish a detection kit for detecting ornithobacterium rhinotracheale hemagglutinin inhibitor antibody. The kit not only can carry out detections for ORT antibodies in chicken serum, but also can carry out detections for yolk antibodies in chicken egg yolks.

Description

technical field [0001] The patent of the present invention relates to a detection kit for the hemagglutination inhibitory antibody of Ornithobacterium rhinotracheale, its preparation method and its application. Wuhan University; preservation date July 6, 2013; strain preservation number is: CCTCC No.: M2013318, referred to as ORT), hemagglutination inhibitory antibody detection kit, preparation method and application, especially for the detection of serum antibodies to Avibacterium gallinarum and semi-quantitative detection kits for egg yolk antibody levels. Background technique [0002] Ornithobacterium rhinotracheale YN12091 (ORT) is a newly named pathogen related to respiratory diseases of poultry and wild birds. The main symptoms of ORT infection are growth retardation, increased mortality, and decreased egg production. Pneumonia, pleurisy and air sacculitis are the main pathological features of this disease. The disease is distributed in the world. ORT may cause primar...

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56911G01N33/86G01N2469/20
Inventor 李富祥李华春廖德芳
Owner YUNNAN ANIMAL SCI & VETERINARY INST
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