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Composition for detecting cardiac troponin I, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method

A technique of cardiac troponin and luminescence immunity, applied in the field of electrochemical detection, can solve the problem of not explaining the mechanism of ischemia, and achieve the effects of short detection time, wide linear range and high sensitivity

Pending Publication Date: 2021-04-13
BEIJING UNIDIAG TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Elevated troponin may indicate myocardial injury but not mechanism of ischemia

Method used

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  • Composition for detecting cardiac troponin I, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method
  • Composition for detecting cardiac troponin I, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method
  • Composition for detecting cardiac troponin I, application of composition, magnetic microsphere electrochemiluminescence immunoassay kit and detection method

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Experimental program
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Effect test

Embodiment 1

[0055] Embodiment 1: the preparation of biotin-labeled cardiac troponin I (Troponin I) antibody and reagent Ra

[0056] Cardiac troponin I (Troponin I) antibody used to label biotin was purchased from Beijing Yuantian Xinye Technology Co., Ltd., the article number is YT-Troponin I-002, and the clone number is 3A1.

[0057] Take 2.0 mg of the antibody used to label biotin cardiac troponin I (Troponin I), use a desalting column PD10 to replace the buffer with phosphate buffer (pH=7.8), use an ultrafiltration tube to concentrate and adjust the concentration to 2.0 mg / mL, add 80 μg of biotin (dissolved in DMF), mix and react for 30 minutes, use desalting column PD10 to remove unlabeled biotin. The biotin-labeled cardiac troponin I (Troponin I) antibody was diluted to 1 mg / L in phosphate buffer (pH=7.4) containing 1% bovine serum albumin, and used as Troponin I reagent Ra. The amount of biotin molecular labeling on the surface of each antibody molecule is 2-3.

Embodiment 2

[0058] Embodiment 2: Preparation of ruthenium-labeled cardiac troponin I (Troponin I) antibody and reagent Rb

[0059] Cardiac troponin I (Troponin I) antibody used to label biotin was purchased from Beijing Yuantian Xinye Technology Co., Ltd., the article number is YT-Troponin I-003, and the clone number is 4F7.

[0060] Take 2.0 mg of the cardiac troponin I (Troponin I) antibody used to label ruthenium terpyridine, use a desalting column PD10 to replace the buffer with phosphate buffer (pH=7.8), use an ultrafiltration tube to concentrate and adjust the concentration to 2.0 mg / mL, add 80 μg of ruthenium succinamide terpyridine (dissolved in DMF), mix and react for 30 minutes, and use desalting column PD10 to remove unlabeled ruthenium. The ruthenium-labeled cardiac troponin I (Troponin I) antibody was diluted to 1 mg / L with 1% bovine serum albumin-containing phosphate buffer (pH=7.4) to serve as Troponin I reagent Rb. The number of ruthenium molecular labels on the surface o...

Embodiment 3

[0061] Embodiment 3: the preparation of calibrator

[0062] The antigen used for the preparation of the calibrator was purchased from Beijing Yuantian Xinye Technology Co., Ltd., and the product number is YT-TroponinI-001. For recombinantly expressed proteins.

[0063] Use 1% bovine serum albumin-containing phosphate buffer (pH=7.4) to dilute the antigen to 5.0 pg / mL, 150 pg / mL according to the marked concentration. Used as a calibrator to establish a standard curve.

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Abstract

The invention belongs to the field of electrochemical detection and relates to to a composition for detecting troponin I, application of the composition, a magnetic microsphere electrochemiluminescence immunoassay kit and a detection method. The method is an electrochemical luminescence method; ruthenium pyridine is adopted as a chemiluminescence marker; the method has the obvious advantages of better sensitivity and better stability; ruthenium is a metal ion, the molecular weight of the ruthenium is small, and the steric hindrance of an antibody is not influenced; the method further has the advantages of short production process, good repeatability, wide detection range, controllable electrochemical luminescence reaction and reduced signal acquisition difficulty.

Description

technical field [0001] The present invention relates to the field of electrochemical detection, in particular to a composition for the detection of cardiac troponin I (Troponin I), the application of the composition, a magnetic microsphere electrochemiluminescence immunoassay kit comprising the composition, and a detection kit based on the composition. The magnetic microsphere electrochemiluminescence immunodetection method of the composition or kit. Background technique [0002] Troponin I (TnI) is a key regulatory protein of striated muscle. Due to its high tissue specificity, cardiac troponin I (cTnI) is a highly sensitive marker of myocardial injury. cTnI can also be used to differentiate skeletal muscle injury (such as rhabdomyolysis, multiple injuries) and myocardial injury. [0003] Elevated cTnI can be seen in acute myocardial infarction (AMI), unstable angina (UAP) and congestive heart failure (CHF). cTnI has been identified as a prognostic marker in patients wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N21/76
CPCG01N33/6887G01N33/6893G01N33/577G01N33/54326G01N21/76G01N2333/4712G01N2800/325G01N2800/10
Inventor 秦军谢元东谢良思黄梦园王晓宁
Owner BEIJING UNIDIAG TECH
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