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Quantitative measurement of cardiac troponin i complex in serum for non-diagnostic purposes

A technique for quantitative measurement of cardiac troponin, applied in the field of quantitative measurement of biological diagnostic markers, can solve problems such as insufficient definition, and achieve the effects of simplifying experimental plans, simplifying preprocessing steps, and eliminating differences in results

Active Publication Date: 2020-01-31
NAT INST OF METROLOGY CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Some standardization problems of biomarkers are insufficient definition of relevant involved entities

Method used

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  • Quantitative measurement of cardiac troponin i complex in serum for non-diagnostic purposes
  • Quantitative measurement of cardiac troponin i complex in serum for non-diagnostic purposes
  • Quantitative measurement of cardiac troponin i complex in serum for non-diagnostic purposes

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Experimental program
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Embodiment Construction

[0033] Main reagents:

[0034] Troponin C: Hytest, Finland;

[0035] Protein precast gel: Solarbio company, China;

[0036] SDS: Beijing Dingguochangsheng Biotechnology Co., Ltd., China;

[0037] Methanol: J.T Baker Company, USA;

[0038] Glacial acetic acid: Beijing Reagent Company, China;

[0039] Glycerin: Sinopharm Group Co., Ltd., China;

[0040] Dithiothreitol DTT: Sigma, USA;

[0041] Tris: Amresco Corporation, USA;

[0042] Bromophenol blue: Sinopharm Group Co., Ltd., China;

[0043] Rainbow pre-stained broad molecular weight protein marker (10-260KD): Zhongke Tairui Company, China;

[0044] Hydrochloric acid: premium grade, Beijing Chemical Reagent Company, China

[0045] Dithiothreitol DTT: Sigma, USA

[0046] Potassium chloride: Sinopharm Group Co., Ltd., China

[0047] Tris: Alfa Aesar, China

[0048] EDTA: Amrescd Corporation, USA

[0049] Calcium chloride: Biodee, China

[0050] Anti-cTnI antibody: 19C7, Hytest, Finland

[0051] Magnetic Beads: Dyna...

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Abstract

The invention discloses a quantitative measurement method of cardiac troponin I compound in serum. The method comprises the following steps: enabling the EDTA to chelate a matrix environment and the Ca2+ contained by the cTnC, and adding the Ca2+ to enable the cTnC and the Ca2+ to recombine, and detecting the Ca2+ through the EDTA chelated Ca2+ and cTnC combined Ca2+, thereby quantitatively measuring all cTnI compound combined with the cTnC. Through the method disclosed by the invention, the measured substance is cleared, the range delimitation of the to-be-measured substance is realized, andthe specific form in the measured component in a mixture is defined. The protein pretreatment step is simplified through an ICP-MS method, other variables imported in the process and the caused uncertainty are eliminated. A detection method of enabling a characteristic peptide fragment to enter mass spectrum is unselected, the result difference caused by the mass-to-change ratio change caused by the heterogeneity is eliminated; recombinant cTnI of the isotope labelling with complex preparation process and difficult representation is unselected as the interior label, and an experimental schemeis simplified; and a standard substance constant value can be finally traced to a SI unit.

Description

technical field [0001] The invention relates to the field of quantitative measurement of biodiagnostic markers, in particular to a method for quantitative measurement of cardiac troponin I complex in serum. Background technique [0002] Cardiac troponin I (cTnI) is currently recognized as a biodiagnostic marker of myocardial infarction because of the excellent correlation between elevated troponin concentrations after myocardial infarction and short-term morbidity and mortality. The accuracy of cardiac troponin I test results directly affects the diagnosis of myocardial infarction and other diseases, and is of great significance for its treatment and prevention. [0003] There are many clinical immunoassay methods for serum stromal cardiac troponin I, but the results are quite different, and the results are affected by the detection methods, antibodies and standard substances. The most accurate reference measurement method for absolute quantification is immunoaffinity enric...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N27/447G01N1/34
CPCG01N1/34G01N27/447G01N33/6848G01N33/6887
Inventor 宋德伟职承瑶李红梅李志林王馨雪肖鹏徐蓓
Owner NAT INST OF METROLOGY CHINA
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