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Test paper card for testing Brucella antibody through competition method

A technology for detection of Brucella bovis and test paper is applied in the field of immunodetection, which can solve the problems of complicated operation steps, difficult to detect antibodies, troublesome operation, etc., and achieve the effects of high sensitivity and repetition rate, easy observation and distinction, and good stability.

Active Publication Date: 2013-06-12
浙江迪恩生物科技股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The isolation and identification of pathogens takes a long time and the operation steps are cumbersome, which is only suitable for the research needs of the laboratory and has no practical value; the complement fixation test is easy to cause hemolysis, and its application is very limited. False positive reaction, and it is difficult to detect the content of the antibody contained in the sample; Tiger red plate agglutination test also has a high false positive reaction, which needs to be improved; ELISA has high sensitivity and good quantitative effect, and is a widely used However, the operation of this method is cumbersome, and the qualitative and quantitative analysis can only be determined by professionals, so it is not suitable for large-scale detection in grassroots and rural areas.

Method used

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  • Test paper card for testing Brucella antibody through competition method
  • Test paper card for testing Brucella antibody through competition method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 Utilizes the sandwich method to detect the detection test paper card structure of the Brucella bovis antibody

[0030] Detection test paper card for detection of bovine Brucella antibody by sandwich method, such as figure 1 and figure 2 As shown, the detection test paper card is composed of a casing 1 and a test strip 2 inside it, and the casing 1 is composed of an upper plate 3 and a lower plate 4, the upper plate 3 has a detection window 5 and a sample injection hole 6, and the test strip 2 It consists of a bottom plate 7 and a sample absorption pad 8 , a colloidal gold marker pad 9 , and a detection reaction area 10 pasted on the bottom plate 7 in sequence. Colloidal gold marker pad 9 is coated with Brucella antigen and colloidal gold marker. The detection reaction area 10 is composed of a test area T coated with Brucella antibodies and a quality control area C coated with Brucella secondary antibodies.

[0031] The sample absorbent pad contains lect...

Embodiment 2

[0033] Embodiment 2 detects the method for bovine brucella antibody

[0034] 1. Preparation of Brucella bovis antigen-colloidal gold marker

[0035] (1) Dialyze the Brucella bovis antigen in 0.005M / L NaCl solution with pH7.0 overnight at 4°C, centrifuge at 10000-12000rpm for 1h at 4°C;

[0036] (2) the pH of the colloidal gold solution of OD526 is adjusted to 4.0-9.0;

[0037] (3) Dilute the Brucella bovis antigen obtained by centrifugation in step (1) to 5 μg / ml~50 μg / ml with 0.005M / L pH9.0 borate buffer solution, and take 1ml of the diluted Brucella bovis antigen Bacterial antigens were added to 1ml colloidal gold solution, oscillated and mixed, and allowed to stand for 5 minutes, then 0.1ml of 10% NaCl solution was added, left to stand for 2 hours after mixing, and centrifuged to obtain colloidal gold-labeled Brucella bovis antigen.

[0038] 2. Processing of blood samples

[0039] Take 100 bovine blood samples immunized with Brucella bovis antigen, collect the blood samp...

Embodiment 3

[0047] Embodiment 3 detects the test paper card false positive rate and the false negative test experiment of bovine brucella antibody

[0048] 1. False Positive Tests

[0049] (1) Take 100 parts of common bovine blood samples, and use a blood pretreatment device to process the blood samples;

[0050] (2) Add 100 μL each of 100 processed common bovine blood samples and 100 μL deionized water into the sample injection hole;

[0051] (2) After the blood sample and deionized water flow through the test area and quality control area of ​​the respective test strips, determine whether the sample contains Brucella bovis antibody. The determination rules are as follows:

[0052] Positive (-): Only a purple-red band appears in the quality control area, and no purple-red band appears in the test area;

[0053] Negative (+): Two purple-red bands appear, one in the test area and the other in the quality control area;

[0054] Invalid: When the quality control area does not display a pu...

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Abstract

The invention discloses a test paper card for testing a Brucella antibody through a competition method and belongs to the technical field of immunodetection. The test paper card is composed of an outer shell and a test paper strip in the outer shell. The outer shell is composed of an upper plate and a lower plate. The upper plate is provided with a testing window and a sample adding hole. The testing paper strip is composed of a base plate and a sample absorption pad, a colloidal gold signing pad and a testing reaction zone which are adhered to the base plate in sequence. The sample absorption pad is aligned to the sample adding hole and the testing reaction zone is aligned to the testing window. The invention further discloses a method for testing the Brucella antibody through the testing paper strip. The Brucella antibody in-site rapid testing paper card is high in sensitivity and repetitive rate, strong in specificity, good in stability, low in fake positive rate and fake negative rate, convenient to use, easy to observe and distinguish and suitable for clinical rapid diagnosis and epidemiological investigation large-scale application in a base level region, countryside and the like.

Description

technical field [0001] The invention belongs to the technical field of immune detection, and in particular relates to a detection test paper card for detecting Brucella bovis antibody by using a competition method. Background technique [0002] Brucella is a Gram-negative short bacillus. When it is first isolated, it is mostly spherical, club-shaped and oval. After subculture, the bacteria gradually become short rod-shaped. The bacteria have no flagella and do not form spores. In 1985, the WHO Expert Committee on Brucellosis divided the genus Brucella into 6 species and 19 biotypes, namely sheep species (biotype 1-3), bovine species (biotype 1-7.9), pig species (biotype 1-7.9), and pig species ( Biotypes 1 to 5) and sheep-type epididymis species, sarira species, and dog species (one biotype each). Clinically, sheep, cattle, and pigs have the greatest significance, and the sheep breed has the strongest pathogenicity. Human disease caused by Brucella is also known by several...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/532
Inventor 张明州王旻子毛开荣陈宗伦吴海芬魏建良程晔
Owner 浙江迪恩生物科技股份有限公司
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