Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Magnetic particle chemiluminiscence detection kit for determining content of hypersensitive troponin T

A technique for supersensitive troponin and troponin, which is applied in the field of magnetic particle chemiluminescence detection kits, can solve the problems of unstable reagents, difficult quantification, and narrow linear range, and achieves improved detection sensitivity, improved stability, and improved specific effect

Pending Publication Date: 2021-03-19
BEIJING LEADMAN BIOCHEM
View PDF6 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods all have certain defects. The RIA method is highly polluted and has gradually withdrawn from the field of clinical testing; the ELISA method is low in automation and complicated to operate, with low sensitivity and narrow linear range; GICA has the advantages of simple operation and fast detection speed, etc. Advantages, but there are also disadvantages such as low sensitivity, unstable reagents, poor repeatability, and difficulty in quantification; CLIA method is an immunoassay technology developed on the basis of enzyme-linked immunoassay, which has high sensitivity and a linear range of detection Wide, simple operation, high degree of automation and other advantages
At present, chemiluminescence immunoassay technology has been widely used because of its many advantages mentioned above, but most of the chemiluminescence immunoassay kits in the prior art are imported closed automatic chemiluminescence detection systems, which require expensive automatic chemiluminescence detectors, thereby limiting the promotion of the use

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
  • Magnetic particle chemiluminiscence detection kit for determining content of hypersensitive troponin T
  • Magnetic particle chemiluminiscence detection kit for determining content of hypersensitive troponin T
  • Magnetic particle chemiluminiscence detection kit for determining content of hypersensitive troponin T

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0060] The preparation method of the magnetic particle chemiluminescence detection kit for measuring the content of hypersensitive troponin T of the present invention comprises the following steps:

[0061] Prepare R1 reagent: 0.9% sodium chloride, the rest is deionized water.

[0062] Prepare the R2 reagent: 1) Prepare the diluent according to the diluent component content of the R2 reagent; 2) Prepare the alkaline phosphatase-labeled troponin T antibody; 3) Use the alkaline phosphatase-labeled troponin T antibody with R2 Reagent diluent for dilution.

[0063] Preparation of magnetic separation reagents: 1) Prepare magnetic particles coated with troponin T monoclonal antibody, measure the magnetic particles, absorb them with a magnet, wash 2 to 3 times with 0.1M Hepes PH7.5, and add the diluent of magnetic separation reagent Make the final concentration of magnetic beads 10mg / ml, add antibody to make the antibody concentration 0.1mg / ml, mix at 37°C for 18 hours, add 10% BSA ...

Embodiment 1

[0079] Example 1 A magnetic particle chemiluminescence detection kit for determining the content of hypersensitive troponin T includes R2 reagent, magnetic separation reagent, calibrator solution series and substrate solution.

[0080] In this embodiment, R1 reagent: 0.9% sodium chloride, and the rest is deionized water.

[0081] R2 reagents include: 1) R2 antibody: alkaline phosphatase-labeled troponin T antibody at a concentration of 0.3 μg / ml; 2) diluent: the diluent of R2 reagent has a pH value of 6.8, including: Tris, at a concentration of 12.04g / L; sodium azide, the concentration is 1g / L; sodium chloride, the concentration is 5.79g / L; bovine serum albumin, the concentration is 10g / L; trehalose concentration is 50g / L; MgCl 2 The concentration is 1.0mM; ZnCl 2 The concentration is 0.1mM; the concentration of Brij 35 is 0.35%; the rest is deionized water.

[0082] In this embodiment, the magnetic separation reagent includes: 1) magnetic particles: magnetic particles coate...

Embodiment 2

[0085] Example 2 Preparation and determination method of the magnetic particle chemiluminescence detection kit for determining the content of hypersensitive troponin T

[0086] (1) First, the 50 μl calibrator series of Example 1 (concentrations are respectively 0, 30, 100, 500, 2000, 6000 pg / ml) were respectively mixed with the 50 μl R1 reagent, 50 μl R2 reagent, and 50 μl magnetic separation reagent of Example 1 in sequence Add to the reaction tube, mix and incubate at 37°C for 10 minutes;

[0087] (2) Wash 3 times with cleaning solution to remove unbound antibodies and impurities;

[0088] (3) Add 150 μl of the luminescence substrate solution in Example 1, and use the Leadman self-developed chemiluminescence detector to measure the relative luminescence intensity (RLU) after the ALP catalyzes the substrate to emit light. The results are shown in the following table:

[0089] Table 1

[0090] Calibrator (pg / ml) 0 30 100 500 2000 6000 RLU(100000) 0.015 ...

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

PropertyMeasurementUnit
Concentrationaaaaaaaaaa
Concentrationaaaaaaaaaa
Concentrationaaaaaaaaaa
Login to View More

Abstract

The invention relates to a magnetic particle chemiluminiscence detection kit for determining the content of hypersensitive troponin T. The magnetic particle chemiluminiscence detection kit comprises areagent R1, a reagent R2, a magnetic separation reagent, a calibrator solution series and a chemiluminiscence substrate solution, wherein the R1 reagent is a buffer solution, and the magnetic separation reagent is a troponin T monoclonal antibody coated magnetic particle solution prepared by dissolving troponin T monoclonal antibody coated magnetic particles in a diluent of the magnetic separation reagent; the reagent R2 is an alkaline phosphatase-labeled troponin T monoclonal antibody solution prepared by dissolving an alkaline phosphatase-labeled troponin T monoclonal antibody in a diluentof the reagent R2, and the chemiluminescent substrate solution is a chemiluminescent substrate solution catalyzed by alkaline phosphatase to emit light. A more accurate, accurate, convenient, rapid and simple method is provided for clinically detecting troponin T in human serum.

Description

technical field [0001] The invention relates to the field of medical diagnostic reagents, in particular to a magnetic particle chemiluminescence detection kit for detecting the content of hypersensitive troponin T by using magnetic particle chemiluminescence technology and antigen-antibody combination technology. Background technique [0002] Troponin is a regulatory protein of muscle contraction and consists of three structurally distinct subunits, troponin T (TnT), troponin I (TnI) and troponin C (TnC). Troponin relaxes actomyosin, allowing actomyosin to interact with myosin to cause muscle contraction. The structure of T and I subunits in the myocardium is different from other muscle tissues. Due to the small molecular weight of cardiac troponin T and I (cTnT, cTnI), the blood concentration increases rapidly after the onset of the disease. cTnT is a polypeptide chain in the molecular structure of striated muscle, with a relative molecular mass of 37×10 3 , there are 4 i...

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
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N21/76
CPCG01N33/6887G01N33/577G01N33/54326G01N21/76
Inventor 巴超杰王鹏杨振军
Owner BEIJING LEADMAN BIOCHEM
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com