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Troponin I competition turbidimetry detecting kit

A detection kit and troponin technology, applied in the medical field, can solve the problems of specificity that is difficult to meet clinical detection requirements, single antibody action site, etc.

Active Publication Date: 2015-10-21
温州煦棠生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the sensitivity in the reagent of measuring troponin I (cTnI) in the above prior art, or the problem that the specificity is difficult to meet the clinical detection requirements, so that the content of troponin I (cTnI) can be clinically determined. Determination standardization; the present invention mixes the sample to be tested with immune polystyrene microspheres, and the troponin I in the sample to be tested reacts with the antibody on the microspheres to undergo an immune complex reaction. Due to the single action site of the antibody, the formation of turbidity is avoided. When the No. 2 reagent is added, the troponin I micro-peptide on the carrier protein undergoes an immune complex reaction with the antibody, and the free antibody concentration obtained by calculating the turbidity intensity can be used to calculate the concentration of the cardiac muscle in the test object. Concentration of calpain I

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] (1) Peptide synthesis: Tribute dual-channel peptide synthesizer (Protein Technologies) was used for solid-phase peptide synthesis.

[0095] (2) Peptide screening: The enzyme-linked immunosorbent overlay experiment was used for peptide screening, and amino acid sequence 8; sequence 9; sequence 10 were selected.

[0096] (3) Carrier protein labeling

[0097] 1) Mix the amino-terminal peptide, central region peptide and carboxy-terminal peptide at 2:3:1 (mass ratio), take 10 mg of the mixed peptide and dissolve it in phosphate buffer with pH=6.5, add 100 mg After EDC was reacted at room temperature for 1 hour, 6 mg of bovine serum albumin was added to shake and react overnight.

[0098] 2) Separating and purifying the reacted bovine serum albumin mixture with a Sephadex G-100 chromatographic column, discarding proteins with a molecular weight lower than 100KD, and retaining cTnI-BSA with a molecular weight above 100KD.

[0099] (4) Immunolatex microsphere labeling

[01...

Embodiment 2

[0111] (1) Peptide synthesis: Tribute dual-channel peptide synthesizer (Protein Technologies) was used for solid-phase peptide synthesis.

[0112] (2) Peptide screening: Peptide screening was performed by enzyme-linked immunosorbent overlay experiment, and sequence 8 was selected.

[0113] (3) Carrier protein labeling

[0114] 1) Dissolve 10 mg of the amino-terminal peptide in phosphate buffer at pH = 6.5, add 100 mg of EDC, react at room temperature for 1 hour, add 6 mg of bovine serum albumin and shake and react overnight.

[0115]2) Separating and purifying the reacted bovine serum albumin mixture with a Sephadex G-100 chromatographic column, discarding proteins with a molecular weight lower than 100KD, and retaining cTnI-BSA-1 with a molecular weight above 100KD.

[0116] (4) Immunolatex microsphere labeling

[0117] Carbodiimide was used to activate the carboxyl group of the microspheres to obtain cTnIAb-PS-1 labeled microspheres with cTnI polyclonal antibody at the ami...

Embodiment 3

[0129] (1) Peptide synthesis: Tribute dual-channel peptide synthesizer (Protein Technologies) was used for solid-phase peptide synthesis.

[0130] (2) Peptide screening: Peptide screening was performed by enzyme-linked immunosorbent immunoassay overlay experiment, and sequence 9 was selected.

[0131] (3) Carrier protein labeling

[0132] 1) Dissolve 10 mg of the peptide in the central region in phosphate buffer at pH = 6.5, add 100 mg of EDC, react at room temperature for 1 hour, then add 6 mg of bovine serum albumin to shake and react overnight.

[0133] 2) Separating and purifying the reacted bovine serum albumin mixture with a Sephadex G-100 chromatographic column, discarding proteins with a molecular weight lower than 100KD, and retaining cTnI-BSA-2 with a molecular weight above 100KD.

[0134] (4) Immunolatex microsphere labeling

[0135] Carbodiimide was used to activate the carboxyl group of the microspheres to obtain cTnIAb-PS-2 labeled microspheres with polyclonal ...

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Abstract

The invention relates to a troponin I competition turbidimetry detecting kit. Three minitype peptide fragments of human cardiac troponin I (cTnI) needed by setting artificial synthesis are used, the peptide fragments and inert carrier protein are coupled, and the peptide fragments and polystyrene latex microballoons prepared by antibodies having an effect on the minitype peptide fragments form reagents. The kit prepared by the immune latex reagents can be used by a semiautomatic or full-automatic developing analyzer for detecting cardiac troponin content in serum.

Description

technical field [0001] The invention relates to a cardiac troponin I competitive immunoturbidimetric assay kit, which belongs to the field of medicine. Background technique [0002] Cardiac troponin I (cTnI) is one of the most sensitive and specific markers of myocardial injury. It is used as the gold standard for judging myocardial cell injury in acute myocardial injury and myocarditis, and also as a risk stratification and prognosis for coronary artery syndrome. key biochemical markers. Troponin is a regulatory protein, the molecule is spherical, and consists of three peptides, troponin T (TnT), troponin I (TnI), and troponin C (TnC), among which troponin I (cTnI) belongs to a A basic protein with a total of 210 amino acids, which only accounts for 4.1% in the free form in serum, most of which are combined with troponin T and C subunits and exist in the form of complexes (Ward DG, Cornes MP, Trayer IP. Structural consequences of cardiac troponin I phosphorylation.J Biol ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 吴蓓蕾李锐唐霞蔚包晓峰
Owner 温州煦棠生物科技有限公司
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