Chemiluminescence immune analytic reagent kit for detecting tuberculosis antibody

A chemiluminescence immunity and chemiluminescence technology, which is applied in the field of immunoanalysis medicine, can solve the problems of time-consuming, low sensitivity and high cost, and achieve the effect of ensuring sensitivity and specificity

Active Publication Date: 2009-03-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The preliminary diagnosis technique of tuberculosis mainly contains: 1, sputum smear examination: simple and easy to do, high accuracy, but the positive rate is low; 2, sputum tuberculosis culture: reliability is high, but time-consuming, expensive and easily affected by medication , the application is limited; 3, chest X-ray: accurate and fast, but easily confused with other lung diseases; the above diagnostic techniques generally have shortcomings such as low sensitivity, poor specificity, and long time consumption, so they have 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

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  • Chemiluminescence immune analytic reagent kit for detecting tuberculosis antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Preparation of the tuberculosis antibody chemiluminescence immunoassay detection kit of the present invention

[0021] The tuberculosis antibody assay kit of the present invention includes: streptavidin-coated microplate, negative and positive controls, biotinylated tuberculosis antigen, alkaline phosphatase-labeled tuberculosis antigen, concentrated washing solution and chemiluminescence Substrate liquid

[0022] 1. Preparation of streptavidin-coated microplate

[0023] (1) Coating

[0024] Mix 0.05M PBS buffer with a pH of 7.2 and appropriate concentration of streptavidin to make a coating solution, add it to the luminescent plate, 100μL per well, overnight at 4°C;

[0025] Specifically, the PBS buffer preparation method is 2.2gNaH 2 PO 4 ·2H 2 O, 12.9gNa 2 HPO 4 ·12H2O, 9gNaCl, dilute to 1000mL with purified water, and adjust the pH to 7.2.

[0026] (2) closed

[0027] Prepare casein blocking solution, respectively add 12.1g Tris, 8.8g NaCl, 10g casein, 1ml Pro...

Embodiment 2～3

[0046] Examples 2 to 3 Preparation of the tuberculosis antibody chemiluminescence immunoassay detection kit of the present invention

[0047] Except that plastic beads and plastic tubes were used as carriers, the other methods were the same as in Example 1 to prepare the tuberculosis antibody chemiluminescence immunoassay detection kit.

Embodiment 4

[0048] Example 4 Method of using the kit of the present invention

[0049] 1) Take out the kit from the refrigerator at 4°C and equilibrate at room temperature for 30 minutes;

[0050] 2) Dilute the concentrated washing solution provided by the kit 25 times with purified water before use;

[0051] 3) Add 50μL of biotinylated antigen to each well; then set up 3 wells of negative control, 50μL per well; 2 wells of positive control, 50μL per well; 1 hole blank, add 50μL of test sample to the remaining wells, use a micro shaker Shake well and mix well, put a sealing film on it, and incubate at 37°C for 30 minutes;

[0052] 4) Wash the plate 5 times, 400μL per well, and finally buckle dry on clean absorbent paper;

[0053] 5) Except for the blank wells, add 50μL of enzyme marker to the remaining wells, shake and mix well, paste the sealing membrane, and incubate at 37°C for 30 minutes;

[0054] 6) Wash the plate 3 times, 400μL per well, and finally buckle dry on clean absorbent paper; ...

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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 present invention relates to the field of immunoassay medicine. Specifically, the present invention uses a combination of chemiluminescence immunoassay and biotin-avidin system to provide a chemiluminescence immunoassay kit for detecting tuberculosis antibodies and a preparation method thereof. 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 throughout the body. Tuberculosis is divided into 5 categories: primary tuberculosis, blood disseminated tuberculosis, secondary tuberculosis, tuberculosis pleurisy and other extra-pulmonary tuberculosis. 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 from AIDS, malaria, diar...

Claims

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Application Information

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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
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