Carbohydrate antigen CA125 quantitative determination kit, and making method and detection method thereof

A technology for CA125 and carbohydrate antigens, applied in the field of quantitative detection of carbohydrate antigen CA125 in blood, can solve the problems of unfavorable automatic detection, unfavorable grass-roots popularization, low detection sensitivity, etc., and achieve favorable signal detection and optimized chemiluminescence enhancement System, the effect of high signal sensitivity

Inactive Publication Date: 2016-03-30
JIANGSU ZECEN BIOTECH CO LTD
View PDF6 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the domestic clinical detection of carbohydrate antigen CA125 is mainly based on 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 CA125 immunochemiluminescence detection kits with independent intellectual property rights are still relatively small, and they only stay at the technical level of 96-well microplates. The detection sensitivity is low, the linearity is narrow, and the specificity is poor, which is not conducive to High-throughput fully automated detection

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
  • Carbohydrate antigen CA125 quantitative determination kit, and making method and detection method thereof
  • Carbohydrate antigen CA125 quantitative determination kit, and making method and detection method thereof
  • Carbohydrate antigen CA125 quantitative determination kit, and making method and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] The configuration of various buffers is as follows:

[0055] 1. Tris salt pH8.0 buffer

[0056] Tris: 12.12mg, sodium chloride 5.82mg, add to 1L of purified water, stir well until completely dissolved, adjust the final pH to 7.5 with hydrochloric acid.

[0057] 2. Preparation of calibrator buffer

[0058] 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.

[0059] 3. Anti-reagent buffer

[0060] 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;

[0061] 4. Magnetic particle buffer

[0062] Tris: 12.12mg, sodium chloride 5.82mg, methyl cellulose ether 50g, add to 1L of purified water, stir well until completely dissolved.

[0063] 5. Luminescence substrate buffer

[0064] Tris12.12g~121.14g, sodium chloride 5.82g, lucigenin 0.03g, a...

Embodiment 2

[0067] Example 2: Preparation of the Quantitative Assay Kit for Carbohydrate Antigen CA125

[0068] 1. Preparation of calibrators and quality controls

[0069] Firstly, the carbohydrate antigen CA125 was dissolved in the standard substance buffer solution, and prepared into calibration products and quality control products with target concentrations as shown in Table 1; the carbohydrate antigen CA125 used in this example was purchased from Fitzgerald, a well-known domestic and foreign manufacturer.

[0070] Table 1: Preparation of calibrators and controls

[0071]

[0072] 2. The preparation method of the anti-reagent is as follows:

[0073] (1), fluorescein isothiocyanate is coupled with carbohydrate antigen CA125 antibody to obtain fluorescein isothiocyanate-labeled CA125 coated antibody:

[0074] First, prepare FITC solution with an anti-reagent buffer solution to a concentration of 2.5 mg / mL FITC, according to the mass ratio of carbohydrate antigen CA125 to FITC as 1:...

Embodiment 3

[0086] Example 3: Steps for detecting carbohydrate antigen CA125 with the carbohydrate antigen CA125 quantitative assay kit

[0087] The method for detecting the carbohydrate antigen CA125 by using the carbohydrate antigen CA125 quantitative assay kit includes the steps of:

[0088] (1) Take three test tubes and add 30 μL carbohydrate antigen CA125 calibrator, 30 μL carbohydrate antigen CA125 quality control, and 30 μL sample to be tested;

[0089] (2) Add 60 μL of anti-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 30 minutes;

[0090] (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 put it in a water bath at 37°C for 5 minutes;

[0091] (4) Precipitate the test tube on the magnetic separator for 2 minutes, slowly invert the test tube and the magnetic separator, pour out the s...

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 discloses a carbohydrate antigen CA125 quantitative determination kit. The kit comprises a calibration substance, a quality control substance, a resisting reagent, a magnetic particle reagent and a luminescence substrate. The invention also discloses a making method of the kit, and a method for detecting a carbohydrate antigen CA125 by using the kit. The resisting reagent is prepared by adopting a fluorescein isothiocyanate labeled CA125 coated antibody and an alkaline phosphatase labeled CA125 labeled antibody, and the magnetic particle reagent is prepared through coupling an anti-fluorescein isothiocyanate antibody with a carboxyl magnetic sphere, so uniform mixing and separation are easy in an immunization reaction, and the reaction speed is greatly improved; and a novel chemiluminescence substrate APLS is adopted as the substrate, so the sensitivity and the specificity of the kit are improved. The determination kit has the advantages of reliable performances, high sensitivity and wide linear range, and can be used together with a semi-automatic and fully-automatic apparatus.

Description

technical field [0001] The invention belongs to the field of biological immune in vitro diagnosis of medical devices, and mainly relates to a kit for quantitatively detecting carbohydrate antigen CA125 in blood based on magnetic particle separation and chemiluminescence method and a preparation method thereof, and the quantitative detection of carbohydrate in blood using the kit Antigen CA125 approach. Background technique [0002] Ovarian malignancy is one of the most malignant gynecological tumors, its incidence rate is second only to breast cancer, and its mortality rate ranks first among gynecological malignancies. Among the primary malignant tumors of the ovary, more than 60% are epithelial ovarian cancer (EOC). Since the ovaries are located in the deep pelvic cavity of women, patients with malignant epithelial ovarian tumors (EOC) generally do not show obvious symptoms and signs in the early stage, and it is difficult to treat them early. It was found that 80% of pati...

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 Applications(China)
IPC IPC(8): G01N33/68G01N33/531G01N33/533G01N21/76
Inventor 夏振伟杨旻汪丹
Owner JIANGSU ZECEN BIOTECH 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