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Carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, preparation method and detection method thereof

A technique for the quantitative determination of carbohydrate antigens, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of unfavorable automatic detection, large economic burden of patients, unfavorable grass-roots popularization, etc., and achieve favorable signal detection and platform stability Long-lasting, easy to mix and separate effects

Inactive Publication Date: 2015-11-25
JIANGSU ZECEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In China, the clinical detection of carbohydrate antigen CA19-9 is mainly based on the application of imported reagents and immunohistochemical reagents. The price of imported reagents is very expensive, which brings a great economic burden to patients and is not conducive to popularization at the grassroots level.
Domestic carbohydrate antigen CA19-9 immunochemiluminescent detection kits are still relatively small, and they only stay at the level of 96-well microplate technology. This technology has low sensitivity, narrow linearity, and poor specificity, which is not conducive to high-throughput Fully automatic detection

Method used

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  • Carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, preparation method and detection method thereof
  • Carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, preparation method and detection method thereof
  • Carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, preparation method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The configuration of various buffers is as follows:

[0053] 1. Tris salt pH8.0 buffer

[0054] Tris: 12.12 mg, sodium chloride 5.82 mg, added to 1 L of purified water, stirred thoroughly until completely dissolved, and adjusted to a final pH of 8.0 with hydrochloric acid.

[0055] 2. Preparation of calibrator buffer

[0056] Add 0.01g of tetracycline and 0.1g of neomycin sulfate to 1L of neonatal bovine serum, fully dissolve and process through a 0.22 μm filter membrane to obtain.

[0057] 3. Anti-reagent buffer

[0058] Tris: 12.12mg~60.57mg, tetracycline: 0.01g~0.05g, sheep serum: 1g~5g, newborn bovine serum 3g~10g, horse serum 1g~5g, add 1L purified water, stir well until completely dissolved;

[0059] 4. Magnetic particle buffer

[0060] Tris: 12.12mg, sodium chloride 5.82mg, methyl cellulose ether 50g, add to 1L of purified water, stir well until completely dissolved.

[0061] 5. Luminescence substrate buffer

[0062] Tris12.12g~121.14g, sodium chloride 5.82...

Embodiment 2

[0065] Example 2: Preparation of the Quantitative Assay Kit for Carbohydrate Antigen CA19-9

[0066] 1. Preparation of calibrators and quality controls

[0067] First: Dissolve the carbohydrate antigen CA19-9 with the standard buffer solution, and prepare the calibrator and quality control product with the target concentration as shown in Table 1;

[0068] Table 1: Preparation of calibrators and controls

[0069]

[0070] 2. The preparation method of the anti-reagent is as follows:

[0071] (1), fluorescein isothiocyanate is coupled with carbohydrate antigen CA19-9 antibody to obtain fluorescein isothiocyanate-labeled CA19-9 coated antibody:

[0072] First, fluorescein isothiocyanate is prepared into a concentration of 2.5 mg / mL fluorescein isothiocyanate solution with anti-reagent buffer, and the ratio of the mass of carbohydrate antigen CA19-9 to fluorescein isothiocyanate is 1: 1.1, transfer the two to a brown glass bottle at the same time, stir at room temperature fo...

Embodiment 3

[0084] Example 3: Steps for detecting the carbohydrate antigen CA19-9 with the carbohydrate antigen CA19-9 quantitative assay kit

[0085] The method for detecting the carbohydrate antigen CA19-9 by using the carbohydrate antigen CA19-9 quantitative assay kit includes the steps of:

[0086] (1) Take three test tubes and add 15 μL carbohydrate antigen CA19-9 calibrator, 15 μL carbohydrate antigen CA19-9 quality control, and 15 μL sample to be tested;

[0087] (2) Add 60 μL of anti-reagent to each test tube, cover the test tube with a plastic film, shake the test tube gently for 30 s, and place it in a water bath at 37°C for 15 minutes;

[0088] (3) Add 30 μL of magnetic particle reagent to each test tube, cover the test tube with plastic film, shake the test tube gently for 30 seconds, and place it in a water bath at 37°C for 5 minutes;

[0089] (4) Precipitate the test tube on the magnetic separator for 2 minutes, slowly invert the test tube and the magnetic separator, pour o...

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Abstract

The invention discloses a carbohydrate antigen 19-9 (CA 19-9) quantitative assay kit, which comprises a CA 19-9 calibration material, a CA 9-19 quality control material, an anti-reagent, a magnetic micro particle reagent, and a luminescent substrate. The invention also discloses a preparation method of the kit and a method using the kit to detect the carbohydrate antigen 19-9 (CA 19-9). Fluorescein isothiocyanate labeled CA 19-9 coated antibody and alkaline phosphatase labeled CA 19-9 labeled antibody are used to prepare the anti-reagent; anti-fluorescein isothiocyanate antibody is coupled to carboxyl magnetic beads to prepare the magnetic micro particle reagent, thus the materials can be more easily mixed and separated in immunity reactions, moreover, the reaction speed is greatly increased; a novel chemical luminescent substrate APLS is taken as the substrate, and the sensitivity and specificity of the kit are both improved. The provided kit has the advantages of high detection sensitivity, good specificity, and little variation, and the shelf life is one year or more.

Description

technical field [0001] The invention belongs to the field of biological immune in vitro diagnosis of medical equipment, and mainly relates to a kit for quantitatively detecting carbohydrate antigen CA19-9 in blood based on magnetic particle separation chemiluminescence method and a preparation method thereof, and the quantitative detection of carbohydrate antigen CA19-9 in blood by using the kit. Method for carbohydrate antigen CA19-9. Background technique [0002] Carbohydrate antigen CA19-9 (Carbohydrate Antigen19-9, CA19-9) is a tumor antigen with very clinical diagnostic value at present. Carbohydrate antigen CA19-9 is a tumor marker related to pancreatic cancer, gallbladder cancer, colon cancer and gastric cancer. Many studies have shown that the content of carbohydrate antigen CA19-9 in tissues is correlated with the development stage of the disease. [0003] In China, the clinical detection of carbohydrate antigen CA19-9 is mainly based on the application of importe...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/531G01N33/533G01N33/535
CPCG01N33/57484G01N33/531G01N33/533
Inventor 杨旻汪丹夏振伟
Owner JIANGSU ZECEN BIOTECH CO LTD
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