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

Luminol chemiluminescence immunological analysis detecting method for cardiac muscle calcium protein

A cardiac troponin and chemiluminescence technology, applied in the field of clinical blood detection and analysis, can solve problems such as difficulties in clinical diagnosis

Inactive Publication Date: 2005-08-31
CHINA PHARM UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the non-tissue specificity of myocardial enzymes and some proteins, it brings difficulties to clinical diagnosis

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] Antibody preparation

[0014] PC89-cTnI 95-108 Transform XL1-Blue competent cells with plasmids and pFD5-cTnI plasmids, pick a single colony and inoculate Amp+Tet double antibody LB culture medium for expansion, add 10 12PFU的VCSM13进行超感染,超感染后的噬菌体用40g / L的PEG 8000及30g / L的NaCl纯化2次,调整噬菌体浓度为0.25mg / ml,加佐剂制成油包水乳剂,免疫新西兰白兔,加强免疫5次后用ELISA方法测定抗体滴度,兔血清用半饱和硫酸铵沉淀粗提,沉淀用适量PBS溶解后,采用Sephedex G-200凝胶层析分离纯化免疫球蛋白G(IgG)。

[0015] 抗体的酶标记

[0016] 辣根过氧化物酶与抗体以1∶1混合,在酸性缓冲液(0.1M MES,0.9M NaCl,0.02%NaN 3 ,pH4.7)中用EDC方法进行连接,反应完成后用Sephedex G-75凝胶层析进行纯化,分部收集,用紫外分光光度计测OD 280 吸光度值,测定各峰辣根过氧化物酶活性及抗体活性,结果显示,辣根过氧化物酶活性峰及抗体活性峰基本重叠,标记结果良好。

[0017] 化学发光免疫分析方法的建立

[0018] 增强剂协同作用发光体系的发光体系条件的优化

[0019] 用正交设计确定最后优化的发光体系:鲁米诺(4.5mmol / L),H 2 o 2 (7.5mmol / L),四苯硼钠(0.8mmol / L),肉桂酸(0.4mmol / L),以pH8.5的0.05mmol / L Tris缓冲液作为缓冲系统。该发光体系发光值比肉桂酸或四苯硼钠单独使用时分别增加2.19和2.05倍,比优化前增加1.18倍,见表1。

[0020] Table1 Effects of different enhancers

[0021] Group number Enhancer n CL(Me...

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 present invention relates to clinical blood detecting and analyzing technology, and is the cTnI Luminol chemiluminescence immunological analysis detecting method with the system including self made cardiac muscle calcium protein resisting antibody I(cTnI) to label horseradish peroxidase, cinnamic acid as luminescence intensifier, tetreaphenyl sodium borate as synergistic intensifier and horseradish peroxidase as catalyst for Luminol-H2O2 luminescence. Compared with available ELISA process, the said CLIA process of the present invention has no obvious difference, correlation coefficient of 0.9852 and diagnosis coincidence rate of 96.15 %.

Description

1. Technical field [0001] The invention belongs to the technical field of clinical blood detection and analysis methods. A quantitative, specific and sensitive method for the clinical detection of cardiac troponin (cTnI) in human serum or plasma was established. A technical platform has been established for matching with the automatic immunoluminescence instrument and the development of other chemiluminescent reagents. 2. Background technology [0002] In recent years, with the changes in people's diet structure and lifestyle, cardiovascular disease has become the primary disease that threatens human health. Therefore, early diagnosis is of great significance to win treatment time and obtain a good prognosis. Acute myocardial infarction (AMI), unstable angina pectoris (UAP), and myocarditis are common clinical cardiovascular diseases. The current laboratory diagnosis mainly includes myoglobin (MA), creatine phosphate Kinase (CK), phosphocreatine kinase isoenzyme (CK-MB), l...

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): G01N21/76G01N33/53G01N33/532
Inventor 沈子龙吴国球
Owner CHINA PHARM 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