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Diagnostic test strip for serological identification of Brucella antibody

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

Active Publication Date: 2019-03-29
洛阳现代生物技术研究院有限公司
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  • Summary
  • 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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  • Diagnostic test strip for serological identification of Brucella antibody
  • Diagnostic test strip for serological identification of Brucella antibody
  • Diagnostic test strip for serological identification of Brucella antibody

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 the preparation of this test strip include LPS antigen and NH antigen. The preparation method is as follows:

[0029] The specific preparation method of LPS antigen: heat the Brucella liquid to 80℃, maintain it for 90min and then inactivate it, sample and inoculate 3 tryptic soy broth solid medium (TSA) plates, each plate is inoculated with 0.1ml, 37 Cultivated for 10 days, all should be aseptically grown. Take the qualified bacterial liquid, centrifuge at 6000 r / min for 30 min, discard the supernatant, and weigh the bacterial pellet. Resuspend each 50g cell pellet in 170ml 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, continue at 66°C Stir for 20 min. Cool to 4°C, centrifuge at 10000r / min for 15min, collect the lower phenol phase and filter it with filter paper, add 500ml of ic...

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] Draw the prepared serum sample with the matching dropper, and add 2~3 drops (about 60μl) vertically and slowly into the sample hole;

[0056] Stand at room temperature for 5~10min to read the result, 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 determined as Brucella infection; when T1 / C value < 0.1 and T2 / C value ≥ 0.1, it is determined as immunopositive; when T1 / C value ≥ 0.1 / C value < 0.1 and T2 / C value < 0.1, judged as negative.

Embodiment 3

[0058] Example 3. Specificity test

[0059]Using Brucella antibody serological differential diagnostic test strips, the positive sera of Yersinia enterica, Brucella crusae, Salmonella Dublin and Escherichia coli 0157 were detected, the T1 / C value and T2 value of 4 specific sera / C values ​​are all within 0.1; T1 / C and T2 / C of Brucella wild virus infection positive serum are 0.964 and 0.145; Brucella S2 vaccine immunopositive serum T1 / C and T2 / C are 0.052 and 1.346; The T1 / C and T2 / C of Brucella-negative serum were 0.030 and 0.062, indicating that the detection method established in this study has strong specificity and no cross-reaction with similar pathogen-positive serum.

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Abstract

The invention relates to a diagnostic test strip for serological identification of Brucella antibody, which belongs to the field of immunology test. The invention comprises a locking shell and a teststrip. The locking shell comprises a locking cover and a locking seat which are fastened to each other. The test strip comprises a bottom plate, and an absorbent pad, a detection pad, a fluorescent microsphere pad 1, a fluorescent microsphere pad 2 and a sample pad which are successively lapped and adhered to the bottom plate. The invention is characterized in that: the test pad is a nitrocellulose membrane provided with a quality control line C, a detection line T1 and a detection line T2; the quality control line C is coated with anti-LPS polyclonal antibodies; the detection line T1 is coated with LPS antigens; the detection line T2 is coated with NH antigens; and the fluorescent microsphere pad 1 and the fluorescent microsphere pad 2 are NH antigens labeled with time-resolved fluorescent microspheres and glass cellulose membranes of LPS antigens respectively. The invention provides a new method for systematically, conveniently and standardizedly detecting the immune antibodies of the S2 vaccine and the infectious antibodies of the disease.

Description

technical field [0001] The invention belongs to the field of immunological detection, in particular to a Brucella antibody serological differential diagnosis test paper. Background technique [0002] Brucellosis (Brucellosis for short) is a zoonotic systemic infectious disease caused by Brucella. Unlike common viral animal diseases, the pathogen is an intracellular parasite , can survive in a variety of livestock. In 1985, the World Health Organization (WHO) divided Brucella into 6 species: sheep, cattle, pigs, dogs and brucella. There are three types of Brucella suis, of which Brucella amphibian 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, and abortion in human beings. In recent years, with the continuous increase of animal husbandry and the frequent circulation of animals and their products in my countr...

Claims

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Application Information

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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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