FPA antibody detection kit for animal brucellosis fluorescence polarization detection method

A technology of brucellosis and fluorescence polarization, applied in the field of zoology, can solve problems such as the initial diagnosis of brucellosis that is not difficult

Inactive Publication Date: 2010-09-29
张晓艳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] According to clinical symptoms, epidemiological characteristics and characteristic lesions, it is not difficult to make a preliminary diagnosis of brucellosis. Presence of similar symptoms necessitates laboratory diagnostic techniques for definitive diagnosis of brucellosis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1, the animal brucellosis fluorescence polarization detection method-FPA antibody detection kit of the present invention, each component is composed as follows:

[0040] (1) Brucella FITC labeled antigen 100ml

[0041] (2) Standard negative serum for animal brucellosis 0.2ml

[0042] (3) Standard positive serum for animal brucellosis 0.2ml

[0043] (4) Standard weak positive serum 0.2ml

[0044] (5) 10 times sample diluent 100ml

[0045] The method for preparing standard negative serum for animal brucellosis is:

[0046] (1) Animals for manufacturing: The cattle for the preparation of positive serum need to undergo laboratory testing. They must be 18-month-old sexually mature, healthy cattle, free of Yarrowia enterocolitica, B. paratyphi, and E. coli O:157 infection. And healthy cattle with negative antibodies such as no reproductive system infection; observe for one week;

[0047] (2) Preparation of immunogen: inoculate Brucella S413 in potato infusion agar flat flasks, in...

Embodiment 2

[0067] Example 2. The method of using the fluorescence polarization detection method for animal brucellosis-FPA antibody detection kit of the present invention is as follows:

[0068] (1) Add 1 ML of 1× sample diluent to a 10×75 mm glass test tube, add 10 μl of standard positive serum or negative serum, and stir three times on a shaker to mix evenly. FPA instrument reads the blank fluorescence density mp value.

[0069] (2) Add 10ul of FITC-labeled antigen with an mp value of 250,000-300,000, mix well, and incubate for 2 minutes at room temperature.

[0070] (3) FPA instrument reads the fluorescence density mp value. If the mp value is higher than the pre-determined threshold 78 to 90mp (the threshold for mp is determined according to different animals, the threshold for sheep and goats is 78mp, the threshold for goats is 88mp, the threshold for pigs is 84mp, the threshold for deer is 87mp, and the threshold for bison is 85mp, the threshold for cattle is 90mp), it is a positive sam...

Embodiment 3

[0076] Example 3, the animal brucellosis fluorescence polarization detection method-FPA antibody detection kit of the present invention was developed as follows: in order to develop a sensitive, specific, and accurate diagnosis of brucellosis, a reagent that satisfies the needs of Chinese animals The need for epidemic prevention, since 2005, Harbin Pinghe Biotechnology Co., Ltd. has cooperated with the Canadian OIE Brucellosis Ground Reference Laboratory to complete the animal brucellosis antibody indirect ELISA rapid detection kit and the basis of the FPA kit. On the previous stage, the production and inspection process research, sensitivity research, specificity research and clinical trials of the FPA test kit were developed, and a draft quality standard for the product was formulated. The relevant issues are explained as follows.

[0077] 1About strain selection and standards

[0078] Strain selection and seed generation. Many Brucella species have been isolated at home and abr...

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Abstract

The invention provides an FPA antibody detection kit for an animal brucellosis fluorescence polarization detection method. The FPA antibody detection kit comprises the following components of 100 ml of brucella FITC labelled antigen, 0.2 ml of animal brucellosis standard negative serum, 0.2 ml of animal brucellosis standard positive serum, 0.2 ml of standard weakly-positive serum and 100 ml of 10-time sample diluent. The FPA antibody detection kit is assembled by using polysaccharide OPS in lipopolysaccharide of smooth type brucellosis as the antigen to mark the fluorescein FITC, preparing positive control serum and standard weakly-positive serum by using inactivated bacterium solution immune healthy cattle or artificially infected healthy cattle, and using healthy non-immune cattle serum as negative control serum and sample diluent. The invention is used for distinguishing mammal brucella infected antibodies and immune antibodies.

Description

(1) Technical field [0001] The invention relates to zoology, in particular to an animal brucellosis fluorescence polarization detection method-FPA antibody detection kit. (2) Background technology [0002] The immunological detection method is a series of experimental methods designed to determine antigens, antibodies, immune cells and the cytokines secreted by immunological theory. With the mutual penetration of disciplines, the scope of immunology has been expanding, and new immunological detection methods have emerged one after another. The scope of application of immunological methods is also expanding day by day. It has not only become an important method for the diagnosis of many clinical diseases, but also provides convenience for the research of many disciplines. [0003] When an antigen meets a corresponding antibody, it can specifically bind, and under the influence of external conditions, a certain reaction phenomenon, such as agglutination or precipitation, can be disp...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/536G01N33/533
Inventor 张晓艳
Owner 张晓艳
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