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A detection test paper card for detecting Brucella bovis antibody by sandwich method

A technology for the detection of Brucella bovis and test strips, applied in the field of immunoassays, can solve the problems of complex processing, cross-reaction, and low sensitivity, and achieve high sensitivity and repeatability, easy to observe the difference, and good stability

Active Publication Date: 2016-05-04
浙江迪恩生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Blood diagnosis can only be used as a preliminary diagnosis method, not as a method of diagnosis; bacteriological examination method takes a long time, the probability of false positive and false negative is high, and the experimental conditions are not easy to control; immunological detection is a rapid detection method, However, the current detection methods all have low sensitivity and cross-reaction; PCR is currently the most commonly used diagnostic method for the disease, but there are complex sample pretreatments, expensive instruments and equipment, time-consuming analysis, and require skilled professional and technical personnel to perform. Completion and other issues
None of these methods are suitable for widespread detection of the disease in grassroots units with simple equipment and lack of technical personnel

Method used

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  • A detection test paper card for detecting Brucella bovis antibody by sandwich method
  • A detection test paper card for detecting Brucella bovis antibody by 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 antibody

[0035] 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 11 coated with a Brucella antigen-carrier protein conjugate and a quality control area 12 coated with a Brucella antibody.

[0036] The carrier protein i...

Embodiment 2

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

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

[0041] (1) Dialyze the Brucella bovis antigen in 0.005M / L pH7.0 NaCl solution at 4°C overnight, 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) The brucella bovis antigen obtained by step (1) centrifugation is diluted to 5 μg / ml~50 μg / ml with the borate buffer solution of 0.005M / LpH9.0, and the brucella bovis after getting 1ml dilution The antigen was added to 1ml of colloidal gold solution, oscillated and mixed, and left to stand for 5 minutes, then 0.1ml of 10% NaCl solution was added, mixed and left to stand for 2 hours, and centrifuged to obtain the colloidal gold-labeled Brucella bovis antigen.

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

[0045] (1) Dissolve 5 mg of Brucella bovis antigen and 2.5 mg of car...

Embodiment 3

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

[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-...

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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 antibody 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 is a Gram-negative short bacillus. When it is first isolated, it is mostly spherical, club-shaped and oval. Brucella can infect a variety of domestic animals and wild animals including humans, mainly through the digestive tract, The skin, mucous membrane, respiratory tract and other ways invade the body, causing similar clinical symptoms after infection, such as long-term fever, miscarriage and infertility, weakness, arthralgia, and hepatosplenomegaly. Bovine brucellos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532
Inventor 王旻子张明洲魏建良程晔陈宗伦吴海芬
Owner 浙江迪恩生物科技股份有限公司
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