Method for detecting cardiac troponin I/T through flash type homogeneous chemiluminescence technology

A cardiac troponin and homogeneous chemiluminescence technology, which is applied in chemiluminescence/bioluminescence, analysis through chemical reactions of materials, biological testing, etc., can solve the problems of cumbersome operation and long detection time, and achieve repeatability Good, less time-consuming, convenient detection process

Pending Publication Date: 2019-07-19
杭州普鲁米生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method uses magnetic particles as a solid support, and requires an external magnetic field and washing steps during the detection process, so it has the disadvantages of cumbersome operation and long detection time

Method used

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  • Method for detecting cardiac troponin I/T through flash type homogeneous chemiluminescence technology
  • Method for detecting cardiac troponin I/T through flash type homogeneous chemiluminescence technology
  • Method for detecting cardiac troponin I/T through flash type homogeneous chemiluminescence technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-1

[0085] Embodiment 1-1, a method for detecting cardiac troponin I by flash-type homogeneous chemiluminescence technology

[0086] cTnI monoclonal antibody Ⅰ is coupled with luminescent substrate 9,10-dihydroacridine, and cTnI monoclonal antibody Ⅱ is coupled with horseradish peroxidase for the detection of cardiac troponin I by flash-type homogeneous chemiluminescent method ; Do the following steps in sequence:

[0087] 1), cTnI monoclonal antibody I coupled with 9,10-dihydroacridine:

[0088] Dissolve 1mg of the luminescent substrate 9,10-acridine in 2ml of DMF to obtain a 9,10-acridine solution;

[0089] Add 100ul of 9,10-acridine solution to 100ul of cTnI monoclonal antibody I with a concentration of 10mg / ml, then add 800ul of 0.05M boric acid buffer (pH8.8), and rotate for 1 hour at room temperature.

[0090] Add the obtained reaction solution into a dialysis bag with a molecular weight cut-off of 7000, put it in PBS buffer (PH=7.4) and dialyze at 4°C for 1 day (24h), dur...

Embodiment 1-2

[0112] Embodiment 1-2, a method for detecting cardiac troponin I by flash-type homogeneous chemiluminescence technology:

[0113] Goat anti-mouse IgG secondary antibody coupled with luminescent substrate 9,10-dihydroacridine, goat anti-rabbit IgG secondary antibody coupled with horseradish peroxidase, used for the detection of cardiac muscle calcium by flash-type homogeneous chemiluminescent method Protein I. Follow the steps in order:

[0114] 1), goat anti-mouse IgG secondary antibody coupled to 9,10-dihydroacridine:

[0115] Dissolve 1mg of the luminescent substrate 9,10-acridine in 2ml of DMF to obtain a 9,10-acridine solution;

[0116] Take 100 ul of 9,10-acridine solution and add 100 ul of goat anti-mouse IgG secondary antibody with a concentration of 10 mg / ml, then add 800 ul of 0.05M boric acid buffer (PH8.8), and rotate at room temperature for 1 hour.

[0117] Add the obtained reaction solution into a dialysis bag with a molecular weight cut-off of 7000, place it i...

Embodiment 2-1

[0156] Example 2-1, a method for detecting cardiac troponin T by flash-type homogeneous chemiluminescence technology

[0157] The cTnT monoclonal antibody Ⅰ is coupled with the luminescent substrate 9,10-dihydroacridine, and the cTnT monoclonal antibody Ⅱ is coupled with horseradish peroxidase for the detection of cardiac troponin T by flash-type homogeneous chemiluminescent method ; Do the following steps in sequence:

[0158] 1), cTnT monoclonal antibody I coupled with 9,10-dihydroacridine:

[0159] Dissolve 1mg of the luminescent substrate 9,10-acridine in 2ml of DMF to obtain a 9,10-acridine solution;

[0160] Take 100 ul of 9,10-acridine solution and add 100 ul of cTnT monoclonal antibody I with a concentration of 10 mg / ml, then add 800 ul of 0.05M boric acid buffer (PH8.8), and rotate for 1 hour at room temperature.

[0161] Add the obtained reaction solution into a dialysis bag with a molecular weight cut-off of 7000, put it in PBS buffer (PH=7.4) and dialyze at 4°C f...

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Abstract

The invention discloses a method for detecting cardiac troponin I / T through a flash type homogeneous chemiluminescence technology. Serum is used as a sample to be detected. The method comprises the steps of: coupling an antibody I to a luminous substrate, so that an antibody I-A is obtained; coupling an antibody II to horse radish peroxidise, so that an antibody II-HRP is obtained; designing two detection systems, adding exciting fluid after incubating the detection system, and instantly detecting a light emitting signal; taking a cTnI / cTnT sample after gradient dilution as a sample, and then,performing detection, so that a formula that the light emitting signal corresponds to the cardiac troponin I / T concentration is obtained; and then, performing detection by taking the sample to be detected as the sample, so that the cardiac troponin I / T concentration in the sample to be detected is finally obtained. The method for detecting the cardiac troponin I / T has the characteristics of beingrapid and high in sensitivity.

Description

technical field [0001] The invention belongs to the biological field, and in particular relates to a method for detecting cardiac troponin I / cardiac troponin T by a flash type homogeneous chemiluminescence method. Background technique [0002] Cardiac troponin is a protein complex that distributes in regular spaces on cardiac tropomyosin and regulates cardiac muscle contraction. It consists of three subunits: cardiac troponin T (cTnT), cardiac troponin I (cTnI) and cardiac troponin C (cTnC). Among them, cTnT and cTnI have unique amino acid sequences and have good myocardial specificity. The content of cTnT and cTnI in normal human serum is very small. When the cardiomyocytes are damaged, the free cTnT and cTnI are released from the cardiomyocytes into the blood rapidly, causing the blood concentration to rise rapidly. The half-life of CTnT is only 2 hours, and the half-life of free cTnI is 2 hours to 5 days. In clinical use, cardiac troponin T (cTnT) is an independent pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/76
CPCG01N21/76G01N33/6887G01N33/6893G01N2800/324
Inventor 江学成杨晓光
Owner 杭州普鲁米生物科技有限公司
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