Cardiac troponin I (cTn I) hypersensitive detection kit and hypersensitive detection method

A technique for cardiac troponin and detection kit, which is applied in biological tests, material inspection products, etc., can solve problems such as insufficient sensitivity for clinical application, inability to accurately detect cTnI, and inability to meet early diagnosis of AMI.

Active Publication Date: 2015-05-20
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the sensitivity of the existing cTnI commercial detection kits is still not high enough for clinical application, and the analytical sensitivity is above 5pg / mL, so it cannot accurately detect ultra-low concentrations of cTnI, so it cannot meet the high requirements of early diagnosis of AMI

Method used

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  • Cardiac troponin I (cTn I) hypersensitive detection kit and hypersensitive detection method
  • Cardiac troponin I (cTn I) hypersensitive detection kit and hypersensitive detection method
  • Cardiac troponin I (cTn I) hypersensitive detection kit and hypersensitive detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Kit 1 was prepared by the following steps:

[0127] Two strains of cTnI antibodies with amino acid segments 18-28 and 83-93 capable of binding to cTnI were used as the first anti-cTnI antibody, and were labeled with ABEI according to the above-mentioned 4 steps of preparation to obtain the first anti-cTnI antibody solution labeled with ABEI. Wherein, the total concentration of the first anti-cTnI antibody is 300ng / mL, and the concentration of ABEI is 30pg / mL.

[0128] Two strains of cTnI antibodies with cTnI-binding amino acid segments 41-49 and 190-196 were used as the second anti-cTnI antibody, and magnetic microspheres were coated according to the above-mentioned steps of Preparation 1 to obtain the second antibody coated with magnetic microspheres. cTnI antibody solution. Wherein, the total concentration of the second anti-cTnI antibody is 5 μg / mL, and the concentration of the magnetic microspheres is 1 mg / mL.

[0129] Dilutions were prepared as in Preparation 11....

Embodiment 2

[0133] Kit 2 was prepared by the following steps:

[0134] Two strains of cTnI antibodies with amino acid segments 26-35 and 83-93 capable of binding to cTnI were used as the first anti-cTnI antibody, and labeled with ABEI according to the above-mentioned 4 steps of preparation to obtain the first anti-cTnI antibody solution labeled with ABEI. Among them, the total concentration of the first anti-cTnI antibody was 300 ng / mL, and the ABEI concentration was 30 pg / mL.

[0135] Two strains of cTnI antibodies with cTnI-binding amino acid segments 41-49 and 169-178 were used as the second anti-cTnI antibody, and a biotinylated second anti-cTnI antibody solution was prepared according to the above-mentioned steps in Preparation 5. Among them, the total concentration of the second anti-cTnI antibody was 300 ng / mL, and the biotin concentration was 30 pg / mL.

[0136] The streptavidin solution coated with magnetic microspheres was prepared according to Preparation 2 above. Wherein, the...

Embodiment 3

[0141] The magnetic microsphere suspension coated with goat anti-FITC polyclonal antibody was prepared according to the above-mentioned preparation 3.

[0142] Using the first anti-cTnI antibody in Example 1, the first anti-cTnI antibody solution labeled with ABEI was prepared according to the above-mentioned Preparation 4.

[0143] Using the second anti-cTnI antibody in Example 1, a FITC solution labeled with the second anti-cTnI antibody was prepared according to Preparation 6 above.

[0144] Dilutions were prepared as in Preparation 11 above.

[0145] The high and low point calibrator solutions were prepared according to Preparation 12 above.

[0146] Each of the solutions prepared above constitutes the components of the kit 3 provided in this example.

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Abstract

The invention provides a cTn I (cardiac troponin I) hypersensitive detection kit and a preparation method thereof. The kit comprises at least one strain of a first anti-cTn I antibody marked with a tracing marker and at least one strain of a second anti-cTn I antibody coated with magnetic micro-spheres, wherein a binding site between the first anti-cTn I antibody and cTn I is different from a binding site between the second anti-cTn I antibody and cTn I; and the kit can preferably further comprise a diluent, and by adopting the diluent, the non-specific binding in a detection process can be significantly reduced, and the detection accuracy and sensitivity can be further improved. The invention also provides a method for detecting cTn I by using the kit, and the method provided by the invention has very high sensitivity, can be used for sensitively and accurately detecting the cTn I content in a sample, and can provide more timely and reliable information for early diagnosis and treatment of AMI.

Description

technical field [0001] The invention relates to the detection field of biochemical substances, in particular to a cardiac troponin I hypersensitive detection kit and a preparation method thereof, and also to a cardiac troponin I hypersensitive detection method. Background technique [0002] Troponin is a regulatory protein of striated muscle contraction, present in cardiac muscle and skeletal muscle, composed of three subunits, namely TnI, TnC and TnT. The three subunits form a complex that plays an important regulatory role in muscle contraction and relaxation. TnI is an inhibitory subunit of actin and has three subtypes: fast skeletal muscle isoform (fTnI), slow skeletal muscle isoform (sTnI) and cardiac muscle isoform (cardiac troponin I, cTnI). Its central cTnI has a molecular mass of 23.9kD and contains 210 amino acid residues. Elevated cTnI content can be used as an important serological indicator for the diagnosis of acute myocardial infarction (AMI). When acute my...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 饶微袁锦云李钦王晟徐红李武李婷华付金秋
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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