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A kind of brucella antibody serological differential diagnosis test paper

A technology for differential diagnosis and Brucella, which is applied in the field of immunology detection, can solve the problems affecting the effect of epidemic prevention work, hindering the culling of brucellosis-infected animals, and being unable to distinguish Brucella infection antibodies from vaccine immune antibodies, etc., to achieve The effect of stable implementation effect, fast method, good sensitivity and specificity

Active Publication Date: 2022-04-05
洛阳现代生物技术研究院有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

LPS and OMPs are the main surface antigens. Although the existing conventional serological detection methods have high sensitivity and specificity, they cannot distinguish Brucella infection antibodies from vaccine immune antibodies, which will affect the effectiveness of epidemic prevention work and hinder Execution of the policy of culling brucellosis-infected animals in my country

Method used

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  • A kind of brucella antibody serological differential diagnosis test paper
  • A kind of brucella antibody serological differential diagnosis test paper
  • A kind of brucella antibody serological differential diagnosis test paper

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, the preparation of test paper

[0028] 1. The main active raw materials for preparing the test paper include LPS antigen and NH antigen, and the preparation method is as follows:

[0029] The specific preparation method of LPS antigen: Heat the Brucella bacterial liquid to 80°C, maintain it for 90 minutes, and then inactivate it. Take samples and inoculate three plates of tryptic soy broth solid medium (TSA), each plate is inoculated with 0.1ml, 37 Cultivate for 10 days at ℃, all should grow aseptically. Take the qualified bacterial solution, centrifuge at 6000r / min for 30min, discard the supernatant, and weigh the bacterial precipitate. Resuspend every 50g of bacterial cell pellet in 170ml of sterilized purified water, heat to 66°C, then add an equal volume of 90% (V / V) phenol solution (water-saturated phenol solution) preheated to 66°C, and continue at 66°C Stir for 20min. Cool to 4°C, centrifuge at 10,000r / min for 15min, collect the lower phenolic ph...

Embodiment 2

[0053] Embodiment 2, the detection method of brucella antibody serological differential diagnosis test paper

[0054] Tear open the aluminum foil packaging bag of the test card, take out the test card, and place it on a flat and clean table;

[0055] Use the matching dropper to draw the prepared serum sample, and drop 2~3 drops (about 60 μl) vertically and slowly into the sample hole;

[0056] Stand at room temperature for 5-10 minutes to read the results, insert the test paper card into the card slot of the fluorescence immunoassay analyzer in the direction of the arrow shown on the card cover, and read the T1 / C value and T2 / C value;

[0057] Judgment criteria: When T2 / C value ≥ 0.1 and T1 / C value ≥ 0.1, it is judged as Brucella infection; when T1 / C value < 0.1 and T2 / C value ≥ 0.1, it is judged as immunopositive; / C value<0.1 and T2 / C value<0.1, judged as negative.

Embodiment 3

[0058] Embodiment 3, specificity test

[0059]Brucella antibody serological differential diagnosis test strips were used to detect the positive sera of Yersinia enterica, Brucella crassa, Salmonella Dublin and Escherichia coli 0157, and the T1 / C values ​​and T2 of the four specific sera / C values ​​were all within 0.1; T1 / C and T2 / C of Brucella wild virus infection positive serum were 0.964 and 0.145; Brucella S2 vaccine immune positive serum T1 / C and T2 / C were 0.052 and 1.346; The T1 / C and T2 / C of Brucella-negative serum were 0.030 and 0.062, which indicated that the detection method established in this study was highly specific and had no cross-reaction with similar pathogen-positive sera.

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Abstract

The invention relates to a brucella antibody serological differential diagnosis test paper, which belongs to the field of immunological detection, and comprises a jamming case and a test paper strip; the jamming case includes a card cover and a card seat which are fastened to each other; the test paper strip includes a bottom plate and sequentially lapped A water-absorbing pad, a detection pad, a fluorescent microsphere pad 1, a fluorescent microsphere pad 2 and a sample pad pasted on the base plate; it is characterized in that: the detection pad is provided with a quality control line C, a detection line T1 and a detection line T2 Nitrocellulose membrane, quality control line C coated with anti-LPS polyantibody, detection line T1 coated with LPS antigen, and detection line T2 coated with NH antigen; the fluorescent microsphere pad 1 and fluorescent microsphere pad 2 are respectively embedding time Glass cellulose membrane for resolving fluorescent microsphere-labeled NH antigen and LPS antigen. The invention provides a new method for systematically, conveniently and standardizedly detecting brucellosis S2 vaccine immune antibody and brucellosis infection antibody.

Description

technical field [0001] The invention belongs to the field of immunological detection, and in particular relates to a serological differential diagnosis test paper for Brucella antibody. Background technique [0002] Brucellosis (brucellosis for short) is a zoonotic systemic infectious disease caused by Brucella. Unlike common viral animal diseases, the pathogen of this disease is an intracellular parasite , can survive in a variety of livestock. In 1985, the World Health Organization (WHO) divided Brucella into 6 species: ovine, bovine, porcine, dog and murine Brucella. There are three types of Brucella suis, of which Brucella melis is the most pathogenic. Brucellosis mainly affects the reproductive organs and joints of cattle and sheep, causing abortion in female animals and orchitis in male animals, as well as fever, joint swelling and pain in humans, and bacterial chronic infectious diseases caused by abortion. In recent years, with the continuous increase in the amoun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/58G01N33/531
CPCG01N33/531G01N33/558G01N33/56911G01N33/582G01N2333/23
Inventor 李秀梅范伟兴李凯尼博狄栋栋原小燕任雪建吴彬刘明瑞杨志肖利侯志乾王路卫一新
Owner 洛阳现代生物技术研究院有限公司
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