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Nanometer magnetic particle chemiluminiscence detection kit for hypersensitive cardiac troponin I and preparation method and application of nanometer magnetic particle chemiluminiscence detection kit

A technology for chemiluminescence detection and cardiac troponin, which is applied in the fields of chemiluminescence/bioluminescence, analysis through chemical reactions of materials, biological testing, etc. It can solve the problems of complex biochemical properties of troponin and inaccurate detection results , to achieve fast and sensitive determination, improved sensitivity, and good system stability

Pending Publication Date: 2022-04-05
HONGKUI BIOLOGICAL CHINA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The performance of cTnI detection reagents is crucial to the judgment of diagnostic results. However, due to the complex biochemical properties of troponin, the development of highly sensitive and specific detection reagents has become a difficult challenge.
A number of research results show that various interference factors such as the influence of autoantibodies, cTnI of various degraded molecular fragments in the patient's blood, etc., these factors may lead to inaccurate test results

Method used

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  • Nanometer magnetic particle chemiluminiscence detection kit for hypersensitive cardiac troponin I and preparation method and application of nanometer magnetic particle chemiluminiscence detection kit
  • Nanometer magnetic particle chemiluminiscence detection kit for hypersensitive cardiac troponin I and preparation method and application of nanometer magnetic particle chemiluminiscence detection kit
  • Nanometer magnetic particle chemiluminiscence detection kit for hypersensitive cardiac troponin I and preparation method and application of nanometer magnetic particle chemiluminiscence detection kit

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preparation example Construction

[0031] In yet another embodiment of the present invention, the preparation method of this hypersensitive cTnI detection kit is provided, comprising the following steps:

[0032] 1. Preparation of M reagent

[0033] 1.1 Preparation of two cTnI-biotin antibodies

[0034] The cTnI monoclonal antibody was labeled with biotin ester, and two kinds of cTnI-biotin antibodies were prepared. The epitopes targeted by the two naked antibodies were different; among them: the antigen targeted by the first human cTnI monoclonal antibody The epitope is Troponin I-Cardiac (41-49Region); the epitope of the second anti-human cTnI monoclonal antibody is Troponin I-Cardiac (87-91Region)

[0035] 1.2 Two cTnI-biotin antibodies were coupled with streptavidin magnetic beads respectively, and the biotinylated cTnI antibody was coated on the streptavidin magnetic beads.

[0036] 1.3 The above reaction was carried out at room temperature, the coating buffer used was Tris buffer, and finally the volume...

Embodiment 1

[0049] Embodiment 1: Nano-magnetic particles chemiluminescent detection kit for ultra-sensitive cardiac troponin I

[0050] 1. Kit composition:

[0051] (1) M reagent: magnetic particles coated with the first anti-human cTnI monoclonal antibody and the second anti-human cTnI monoclonal antibody; Tris buffer.

[0052] (2) R reagent: alkaline phosphatase (AP) labeled third anti-human cTnI monoclonal antibody; Tris buffer.

[0053] 2. Preparation of the kit:

[0054] (1) The first anti-human cTnI monoclonal antibody and the second anti-human cTnI monoclonal antibody were respectively subjected to a labeling reaction and coupled with biotin to obtain biotinylated first anti-human cTnI monoclonal antibody and biotinylated Second anti-human cTnI monoclonal antibody; then biotinylated first anti-human cTnI monoclonal antibody and biotinylated second anti-human cTnI monoclonal antibody and streptavidin-modified magnetic particles were placed at room temperature React on a mixer for...

Embodiment 2

[0056] Example 2: Methodological investigation of nano-magnetic particle chemiluminescent detection kit for ultra-sensitive cardiac troponin I

[0057] 1. Minimum detection limit

[0058] Use the zero-concentration calibrator as a sample for detection, repeat the measurement 20 times, obtain the RLU value of the 20 measurement results, and calculate the average value and standard deviation (SD). inferred According to the concentration-chemiluminescence (RLU) value results between the zero-concentration calibrator and the adjacent calibrator, a two-point regression fitting is carried out to obtain a linear equation, which is Put the corresponding RLU value into the equation to find the corresponding concentration value, the results are shown in Table 1:

[0059] Table 1: Determination results of the minimum detection limit

[0060]

[0061]

[0062] The above results show that the minimum detection limit of the nano-magnetic particle chemiluminescent detection kit ...

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Abstract

The invention discloses a nano magnetic particle chemiluminiscence detection kit for hypersensitive cardiac troponin I as well as a preparation method and application of the nano magnetic particle chemiluminiscence detection kit. The detection kit comprises: magnetic particles coated with a first anti-human cTnI monoclonal antibody and a second anti-human cTnI monoclonal antibody; the third anti-human cTnI monoclonal antibody is labeled by alkaline phosphatase; the first anti-human cTnI monoclonal antibody and the second anti-human cTnI monoclonal antibody are respectively specific to different antigen epitopes of human cTnI. The nano magnetic particle chemiluminescence detection kit has the characteristics of high detection sensitivity, good specificity and good stability. Compared with a conventional detection reagent for detecting troponin I, the troponin I detection reagent has great advantages and can meet the requirements of clinical application.

Description

technical field [0001] The invention relates to the technical field of medical diagnostic reagents, in particular to a nano-magnetic particle chemiluminescent detection kit for hypersensitive cardiac troponin I, a preparation method and application thereof. Background technique [0002] Troponin is composed of T, C, and I subunits, and together with tropomyosin, regulates the interaction between actin and myosin by regulating the activity of calcium ions on striated actin ATPase. When the myocardium is damaged, the cardiac troponin complex is released into the blood, and after 4-6 hours, it begins to rise in the blood, and the highest value appears at 12-24 hours. Increased cardiac troponin can still be detected in peripheral blood 7-10 days after myocardial injury. [0003] There are three isomeric forms of troponin, and cardiac troponin I (cTnI) is only present in cardiac tissue. The specific detection of cTnI can be realized by using anti-cTnI monoclonal antibody, so cT...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N33/58G01N21/76
Inventor 王保全
Owner HONGKUI BIOLOGICAL CHINA CO LTD
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