Test paper card for testing Brucella antibody through sandwich method

A technology for detection of Brucella bovis and test paper, applied in the field of immunodetection, can solve the problems of high false positive rate, lack of technical personnel, long reaction time, etc., and achieve high sensitivity and repetition rate, easy to observe and distinguish, and good stability. Effect

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

AI Technical Summary

Problems solved by technology

[0005] In my country, the standard method for the detection of brucellosis is WS269-2007, but its sensitivity and specificity are poor, the reaction time is long, and the false positive rate is relatively high. It is a technology that has been eliminated in international trade.
Enzyme-linked immunosorbent assay (ELISA) and PCR are commonly used diagnostic methods for this disease, but they all have problems such as complicated sample pretreatment, expensive instruments and equipment, time-consuming analysis, and requiring skilled professional and technical personnel to complete.
However, brucellosis occurs mostly in rural areas and pastoral areas, and grassroots units, especially pastoral areas, have simple equipment and lack of technical personnel, making it difficult to carry out extensive detection of the disease

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 Utilizes the sandwich method to detect the detection test paper card structure of the Brucella bovis antigen

[0035] Detection test paper card for detecting Brucella bovis antigen 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 antibody and colloidal gold marker. The detection reaction area 10 is composed of a test area 11 coated with a Brucella antibody-carrier protein conjugate and a quality control area 12 coated with a Brucella antigen.

[0036] The carrier protein is bov...

Embodiment 2

[0039] Embodiment 2 detects the method for bovine brucella antigen

[0040] 1. Preparation of Brucella bovis antibody-colloidal gold marker

[0041] (1) Dialyze the Brucella bovis antibody in 0.005M / L NaCl solution with pH7.0 overnight at 4°C, centrifuge at 10000-12000rpm for 1h at 4°C;

[0042] (2) the pH of the colloidal gold solution of OD526 is adjusted to 4.0-9.0;

[0043] (3) Dilute the Brucella bovis antibody obtained by centrifugation in step (1) with 0.005M / L pH9.0 borate buffer solution to 5 μg / ml~50 μg / ml, and take 1ml of the diluted Brucella bovis antibody Bacteria antibody was added to 1ml colloidal gold solution, oscillated and mixed, and allowed to stand for 5 minutes, then added 0.1ml of 10% NaCl solution, mixed and left to stand for 2 hours, centrifuged and purified to obtain colloidal gold-labeled Brucella bovis antibody.

[0044] 2. Preparation of Brucella bovis antibody-carrier protein conjugate

[0045](1) Dissolve 5 mg of Brucella bovis antibody and 2....

Embodiment 3

[0057] Embodiment 3 detects the test strip false positive rate and the false negative test experiment of bovine brucella antigen

[0058] 1. False Positive Tests

[0059] (1) Take 100 parts of common bovine blood samples, and use a blood pretreatment device to process the blood samples;

[0060] (2) Add 100 μL each of 100 processed common bovine blood samples and 100 μL deionized water into the sample injection hole;

[0061] (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:

[0062] Negative (-): Only a purple-red band appears in the quality control area, and no purple-red band appears in the test area;

[0063] Positive (+): Two purple-red bands appear, one in the test area and the other in the quality control area;

[0064] Invalid: When the quality control area does not display a purple-r...

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Abstract

The invention discloses a test paper card for testing a Brucella antibody through a sandwich 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 antigen by using a sandwich method. Background technique [0002] Brucellosis (Brucellosis) is a kind of zoonotic systemic infectious disease caused by Brucella, which is widely prevalent all over the world. It is a second-class animal disease and an infectious disease that must be inspected in international trade inspections. Brucella can infect a variety of livestock and wild animals, including humans, and invade the body mainly through the digestive tract, skin mucous membrane, respiratory tract, etc., causing similar clinical symptoms after infection, such as long-term fever, abortion and infertility, weakness , joint pain and hepatosplenomegaly. In 1897, the Danish doctor Bang discovered Brucella abortus, also known as Brucella abortus, which caused cattle abortion. Brucellosis exists widely in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532
Inventor 张明洲王旻子程晔魏建良吴海芬陈宗伦
Owner 浙江迪恩生物科技股份有限公司
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