Chemiluminescence immune analytic reagent kit for detecting tuberculosis antibody

A technology of chemiluminescent immunity and tuberculosis antibody, which is applied in the field of immunoanalysis medicine to achieve the effect of ensuring sensitivity and specificity

Active Publication Date: 2013-04-03
CHEMCLIN DIAGNOSTICS CO LTD
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The primary diagnostic techniques for tuberculosis mainly include: 1. Sputum smear examination: simple and easy to do, high accuracy, but low positive rate; 2. Sputum tuberculosis culture: High reliability, but time-consuming, expensive, and easily affected by medication, the application is limited; 3. Chest X-ray: accurate and fast, but easy to be confused with other lung diseases; the above diagnostic techniques generally have low sensitivity, poor specificity, and consumption Shortcomings such as length of time, so it has limited diagnostic value for tuberculosis
With the rapid development of molecular biology technology, highly sensitive methods are used to detect tuberculosis and its specific DNA fragments, such as PCR, biological probes and gene chips, etc., which have the advantages of simplicity, sensitivity and specificity in the detection of tuberculosis. However, corresponding testing equipment is required, the technical requirements are high, the cost is high, and it is difficult to promote
At present, ELISA, gold standard and other immunodiagnostic techniques for detecting tuberculosis antibodies in serum are simple and fast, do not require sophisticated instruments, and are easy to promote. However, the currently used diagnostic antigens have the disadvantages of high false positive rate and poor specificity. Antigenous and immunogenic diagnostic antigens, combined detection of several antigens can improve sensitivity without affecting specificity, and has higher sensitivity than single antigen method, which is of great significance to the diagnosis and methodological establishment of tuberculosis

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Chemiluminescence immune analytic reagent kit for detecting tuberculosis antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Preparation of tuberculosis antibody chemiluminescence immunoassay detection kit of the present invention

[0022] The tuberculosis antibody assay kit of the present invention comprises: streptavidin-coated microwell plate, negative and positive controls, biotinylated tuberculosis antigen, alkaline phosphatase-labeled tuberculosis antigen, concentrated washing solution and chemiluminescence substrate solution;

[0023] 1. Preparation of streptavidin-coated microwell plates

[0024] (1) Coated

[0025] Mix 0.05M PBS buffer solution with a pH value of 7.2 and appropriate concentration of streptavidin to make a coating solution, add it to the luminescence plate, 100 μL per well, and overnight at 4 °C;

[0026] Specifically, the PBS buffer preparation method is 2.2gNaH 2 PO 4 2H 2 O, 12.9gNa 2 HPO 4 12H 2 O, 9gNaCl, dilute to 1000mL with purified water, adjust pH to 7.2.

[0027] (2) closed

[0028] Prepare casein blocking solution, respectively add 12.1...

Embodiment 2～3

[0047] Embodiment 2~3 preparation tuberculosis antibody chemiluminescence immunoassay detection kit of the present invention

[0048] Except for using plastic beads and plastic tubes as carriers, the tuberculosis antibody chemiluminescence immunoassay detection kit was prepared in the same manner as in Example 1.

Embodiment 4

[0049] Embodiment 4 The using method of kit of the present invention

[0050] 1) Take out the kit from the refrigerator at 4°C, and equilibrate at room temperature for 30 minutes;

[0051] 2) Dilute the concentrated washing solution provided by the kit 25 times with purified water before use;

[0052] 3) Add 50 μL of biotinylated antigen to each well; then set 3 wells for negative control, 50 μL for each well; 2 wells for positive control, 50 μL for each well; 1 well for blank, add 50 μL of sample to be tested in the remaining wells, and use a micro shaker Shake and mix well, affix the sealing film, and incubate at 37°C for 30 minutes;

[0053] 4) Wash the plate 5 times, 400 μL per well, and finally dry it on clean absorbent paper;

[0054] 5) Add 50 μL of enzyme markers to the remaining wells except for the blank well, vortex and mix well, stick the sealing film on the plate, and incubate at 37°C for 30 minutes;

[0055] 6) Wash the plate 3 times, 400 μL per well, and fina...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a reagent kit for detecting tuberculosis antibodies, and a preparation method thereof. The kit is composed of a chain avidin coated solid-phase vector, negative and positive controls, biotinylation tuberculosis antigens, alkaline phosphatase marker tuberculosis antigens, concentration washing solutions and chemiluminescent substrate solutions. The reagent kit adopts the double antigen sandwich reaction principle and the biotin-avidin system coated solid-phase vectors, the method not only has uniform and firm coating and greatly improves the antigen purity, but also can increase the antigen absorption, lower the cost and have the characteristics of high sensitivity, strong specificity and the like, can accurately detect the tubercubosis antibodies in various tuberculosis serums, and provide a reliable basis for clinical diagnosis and tuberculosis screening.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a chemiluminescence immunoassay kit for detecting tuberculosis antibodies and a preparation method thereof by combining chemiluminescence immunoassay with a biotin-avidin system. Background technique [0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis. Tuberculosis, also known as tuberculosis and "white plague", is an ancient infectious disease that can invade various organs of the human body. Tuberculosis is divided into 5 categories: primary pulmonary tuberculosis, hematogenously disseminated pulmonary tuberculosis, secondary pulmonary tuberculosis, tuberculous pleurisy and other extrapulmonary tuberculosis, etc. Tuberculosis is one of the most threatening infectious diseases to humans today, and it is also a single The most common cause of death caused by infectious agents has exceeded the total number of deaths...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/543G01N21/76
Inventor 赵娟娟应希堂宋胜利胡国茂郑金来张坤
Owner CHEMCLIN DIAGNOSTICS CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap