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

Reagent kit

A kit and plate-type technology, applied in the field of enzyme-labeled immunoassay technology, can solve the problems of expensive, difficult to be widely used in clinical practice, and the sensitivity of general enzyme immunoassay methods cannot meet the requirements, and achieve low cost and low negative test results. bottom effect

Inactive Publication Date: 2003-03-12
SHANGHAI SECOND MEDICAL UNIV
View PDF0 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Corresponding imported kits are used in China, which are expensive and difficult to be widely used in clinical practice
[0007] Under normal circumstances, the content of cardiac troponin I in the blood is very low (<0.1 μg / L), and the sensitivity of general enzyme immunoassay methods cannot meet this requirement

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
  • Reagent kit
  • Reagent kit
  • Reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 An antibody, antigen or immune complex was indirectly immobilized on a solid phase by a streptavidin-biotin capture system, and a plate-type streptavidin two-site sandwich one-step method was established. Among them, the plate-type streptavidin solid-phase two-site sandwich one-step method is to pre-coat the enzyme label plate with streptavidin, biotin-labeled anti-cTnI monoclonal antibody, and horseradish peroxidase to label another anti-cTnI antibody. cTnI monoclonal antibody.

[0020] Preparation of streptavidin solid phase: take 50μl streptavidin (1mg / ml), add 200μl distilled water to dilute, add 50μl 3mol / l acetic acid, leave at room temperature for 5min, adjust to pH 6.0 with 2mol / l Tris, and then use Coating buffer (0.05mol / l, pH 9.5 carbonate buffer) was diluted to 2μg / ml, added to polystyrene ELISA microplate, 120μl / well at 4°C overnight, washed 3 times with washing solution, added blocking solution (0.5 %BSA, 0.02mol / l, pH 7.2 PBS) at 37°C for 1 hou...

Embodiment 2

[0024] Two-site sandwich one-step ELISA method for quantitative detection of serum cTnI

[0025]Figure 1 is the operation flow chart of the plate-type streptavidin solid-phase two-site sandwich one-step ELISA for quantitative detection of serum cTnI created by the present invention. Or hold the test serum sample, then add 100 μl of biotinylated anti-TnI monoclonal antibody (about 2 μg / ml) and a mixture of horseradish peroxidase-labeled anti-TnI monoclonal antibody (pH 7.2, 0.02mol / l phosphate buffer). solution), react at 37°C for 90 minutes, and wash. Add o-phenylenediamine (OPD) substrate solution at 37° C. for 15 minutes, and add 2 mol / l sulfuric acid to stop the reaction. The absorbance value (A) at 490nm was measured on a microplate reader, with A490nm as the ordinate and each TnI concentration as the abscissa, and a standard curve was drawn.

[0026] Methodological examination:

[0027] Standard curve: use purified bovine cardiac muscle TnI antigen, after quantificatio...

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

A reagent kit for enzyme-linked immunodetection is characterized by that on the basis of ELISA method, the chain avidin-biotin capture system is used to indirectly fix the antibody, antigen, or immune complex to solid phase and the plate-type chain avidin dual-site sandwich one-step ELISA for quantitatively detecting the serum cTnI. Its advantages are simple method, high speed, and low-negative background.

Description

technical field [0001] The kit of the invention relates to biological technology, in particular to an enzyme-labeled immunoassay technology. The streptavidin and biotin capture technology is applied to the enzyme-linked immunology technology, and a plate-type streptavidin double-site sandwich one-step quantitative enzyme method is established. Streptavidin-ELISA was used to detect serum cTnI levels in patients. Background technique [0002] In my country, with the development of the national economy and the improvement of people's living standards, the morbidity and mortality of cardiovascular diseases are increasing year by year, so it is necessary to strengthen the diagnosis of early AMI. The diagnosis of ischemic heart disease is divided into three aspects: biochemical examination, ECG changes and persistent chest pain. In biochemical tests, the indicators considered to be effective in diagnosing myocardial ischemic injury are: troponin T (TnT) and I (TnI). TnT, TnI and...

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): C12Q1/02G01N33/53G01N33/535G01N33/543G01N33/577
Inventor 郑佐娅王鸿利
Owner SHANGHAI SECOND MEDICAL UNIV
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