Immunological kit for detecting florfenicol

The technology of florfenicol and reagent kit is applied in the field of immunological reagent kit for detecting florfenicol, which can solve the problems of complex and expensive instruments, professional personnel required for operation, and high detection cost, and achieve wide application prospects, low price, The effect of high sensitivity

Active Publication Date: 2018-06-15
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the extensive and non-standard use of florfenicol, its residues in animal foods such as livestock, poultry, and aquatic products have also been paid more and more attention.
[0003] At present, the main method for detecting florfenicol in animal foods is the instrument method. Although the detect

Method used

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  • Immunological kit for detecting florfenicol
  • Immunological kit for detecting florfenicol
  • Immunological kit for detecting florfenicol

Examples

Experimental program
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Effect test

Embodiment 1

[0064] Embodiment 1 florfenicol complete antigen

[0065] Florfenicol complete antigen is a conjugate of a Florfenicol derivative and a carrier protein, wherein the Florfenicol derivative is abbreviated as FFOA, and its chemical formula is as follows:

[0066]

[0067] The carrier protein is bovine serum albumin (BSA) or chicken ovalbumin (OVA).

[0068] The obtained complete antigen FFOA-BSA is used as the immunogen for the preparation of the camelid antibody, and the complete antigen FFOA-OVA is used as the coating agent constructed by the florfenicol enzyme-linked immunoassay method.

Embodiment 2

[0069] Example 2 Preparation of Florfenicol Alpaca Polyclonal Antibody

[0070] (1) Mix the florfenicol complete antigen FFOA-BSA and Freund's complete adjuvant in equal proportions and fully emulsify the alpaca by subcutaneous injection in the neck and back at a dosage of 750 μg / time;

[0071] (2) Boost immunization on days 22, 36, 50, and 64 respectively (mix FFOA-BSA and Freund's incomplete adjuvant in equal proportions and fully emulsify them, and immunize alpacas by subcutaneous injection at multiple points on the back, with a dosage of 500 μg / time), a total of 4 booster immunizations, with an interval of 2 weeks.

[0072] (3) On the 71st day, 50 mL of whole blood was collected from the neck of the alpaca, placed in a water bath at 37°C for 30 minutes, centrifuged at 12,000 rpm at 4°C for 30 minutes, and the supernatant collected was Florfenicol Alpaca Cloned antibody, aliquoted and stored at -20°C.

[0073] The alpaca was immunized with florfenicol complete antigen F...

Embodiment 3

[0074] Example 3 Preparation of Florfenicol Bactrian camel polyclonal antibody

[0075] (1) The florfenicol complete antigen FFOA-BSA and Freund's complete adjuvant were mixed in equal proportions and fully emulsified to immunize Bactrian camels by multi-point injection in the neck and back, with a dose of 500 μg / time;

[0076] (2) Immunization was boosted on days 15, 29, 43, 57, and 71 (mix FFOA-BSA with Freund's incomplete adjuvant in equal proportions and fully emulsify them, and immunize both sides by subcutaneous injection at multiple points on the neck and back). humpback camel, the dosage is 500μg / time), a total of 5 times of booster immunization, the immunization interval is 2 weeks.

[0077] (3) On the 78th day, 100 mL of whole blood was collected from the neck of the Bactrian camel, placed in a water bath at 37°C for 30 minutes, centrifuged at 12,000 rpm at 4°C for 30 minutes, and the supernatant collected was the florfenicol doublet Camel polyclonal antibody, aliqu...

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Abstract

The invention discloses an immunological kit for detecting florfenicol. A florfenicol immunodetection antibody is prepared by immunizing a Camelidae animal with a conjugate of a florfenicol derivativeand a carrier protein as a florfenicol complete antigen. The high-affinity florfenicol Camelidae animal antibody is obtained in the invention, the alpaca polyclonal antibody titer of florfenicol is 1:32000, and the Bactrian camel polyclonal antibody titer of florfenicol is 1:256000; and a florfenicol enzyme-linked immunoassay kit can be developed in the invention, the florfenicol detection limitof the kit is 0.3 ng/mL, the half-inhibitory concentration is 13.3 ng/mL, and the kit can achieve high-flux, fast and highly-sensitive florfenicol detection requirements. The kit can be used for the residual detection of florfenicol in livestock and poultry, aquatic products and other animal foods, has the advantages of easiness in operation, high in sensitivity and low price, is very suitable forscreening a large number of samples, and has wide application prospects.

Description

technical field [0001] The invention relates to the technical field of food safety immunoassay detection technology, and more specifically relates to an immunology kit for detecting florfenicol. Background technique [0002] Since Announcement No. 193 of the Ministry of Agriculture in 2002 clearly stipulated that chloramphenicol should be banned from all food animals, florfenicol has become the main substitute after the ban on chloramphenicol. Florfenicol has strong bactericidal power and low drug resistance rate after use. It is widely used in bacterial diseases of livestock, poultry and aquatic animals, and has a good therapeutic effect. However, with the extensive and non-standard use of florfenicol, its residues in animal foods such as livestock, poultry, and aquatic products have been paid more and more attention. [0003] At present, the main method for detecting florfenicol in animal foods is the instrument method. Although the detection of florfenicol in the instrum...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/535G01N21/31G01N33/02
CPCG01N21/31G01N33/02G01N33/535G01N33/54306
Inventor 王弘王锋沈玉栋张瑾如杨金易李然梁一凡肖治理
Owner SOUTH CHINA AGRI UNIV
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