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High-sensitivity magnetic particle chemiluminescence immunoassay kit for cardiac troponin I (hs-cTnI) and preparation method and purpose thereof

A technology of cardiac troponin and magnetic particles, which is applied in the field of medical testing, can solve problems such as the inability to detect changes in cTnI concentration, and achieve the effects of stable test results, wide detection range, and high sensitivity

Inactive Publication Date: 2019-05-03
SHENZHEN AMTECH BIOENGINEERING LTD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional cTnI chemiluminescent immunoassays cannot detect changes in cTnI concentration within 4-6 hours after the onset of myocardial infarction due to the limitation of analytical sensitivity, and it usually takes 7 hours to make a diagnosis

Method used

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  • High-sensitivity magnetic particle chemiluminescence immunoassay kit for cardiac troponin I (hs-cTnI) and preparation method and purpose thereof
  • High-sensitivity magnetic particle chemiluminescence immunoassay kit for cardiac troponin I (hs-cTnI) and preparation method and purpose thereof
  • High-sensitivity magnetic particle chemiluminescence immunoassay kit for cardiac troponin I (hs-cTnI) and preparation method and purpose thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1——Preparation of high-sensitivity cardiac troponin I (hs-cTnI) magnetic particle chemiluminescence immunoassay kit

[0035] 1. Preparation of Reagent R1

[0036] A. Preparation of biotin-labeled anti-cTnI polyclonal antibody: Dilute anti-cTnI polyclonal antibody (HyTest) with buffer to a concentration of 2 mg / mL, mix with biotin and react at room temperature for 1-3 hours for biotin labeling . The mass ratio of anti-cTnI polyclonal antibody to biotin is 5:1-20:1, wherein the biotin is excessive, so as to fully label the anti-cTnI polyclonal antibody. The composition of the buffer is 5-50mM Na 2 CO 3 +10-100mM NaHCO 3 , pH 8.0-10.5. After labeling, use a Thermo Scientific Zeba TM Spin Desalting Columns were subjected to centrifugal desalting and purification to obtain biotin-labeled anti-cTnI polyclonal antibody, which was stored at 4°C for future use.

[0037] B. Prepare the prepared biotin-labeled anti-cTnI polyclonal antibody with capture antibody dil...

Embodiment 2

[0052] Example 2——Preparation of high-sensitivity cardiac troponin I (hs-cTnI) magnetic particle chemiluminescence immunoassay kit

[0053] The preparation method of the present embodiment is similar to Example 1, but has the following differences:

[0054] 1. Preparation of Reagent R1

[0055] The prepared biotin-labeled anti-cTnI polyclonal antibody was adjusted to a concentration of 0.4 μg / mL with capture antibody diluent. The composition of the capture antibody diluent is shown in Table 4 below.

[0056] The composition of the capture antibody diluent of table 4 embodiment 2

[0057]

[0058] 2. Preparation of Reagent R2

[0059] The prepared two strains of acridinium ester-labeled anti-cTnI monoclonal antibodies targeting different cTnI epitopes were mixed at a mass ratio of 1:1, and then prepared to a concentration of 0.5 μg / mL with labeled antibody diluent. The composition of the labeled antibody diluent is shown in Table 5 below.

[0060] The composition of the...

Embodiment 3

[0067] Example 3——Preparation of high-sensitivity cardiac troponin I (hs-cTnI) magnetic particle chemiluminescence immunoassay kit

[0068] The preparation method of the present embodiment is similar to Example 1, but has the following differences:

[0069] 1. Preparation of Reagent R1

[0070] The prepared biotin-labeled anti-cTnI polyclonal antibody was adjusted to a concentration of 2 μg / mL with capture antibody diluent. The composition of the capture antibody diluent is shown in Table 7 below.

[0071] The composition of the capture antibody diluent of table 7 embodiment 3

[0072]

[0073] 2. Preparation of Reagent R2

[0074] The prepared two strains of acridinium ester-labeled anti-cTnI monoclonal antibodies targeting different cTnI epitopes were mixed at a mass ratio of 1:1, and then prepared at a concentration of 5 μg / mL with labeled antibody diluent. The composition of the labeled antibody diluent is shown in Table 8 below.

[0075] The composition of the lab...

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Abstract

The application provides a high-sensitivity magnetic particle chemiluminescence immunoassay kit for cardiac troponin I (hs-cTnI). The kit comprises a reagent R1, a reagent R2, a magnetic particle working solution and a cTnI antigen calibrator. The reagent R1 contains a biotin-labeled anti-cTnI polyclonal antibody; the reagent R2 contains two acridinium-ester-labeled anti-cTnI monoclonal antibodiesagainst different cTnI epitopes; the magnetic particle working solution includes streptavidin magnetic beads. In addition, the kit also can include a H2O2 solution and an alkaline solution. Besides,the application also provides a preparation method and purpose of the kit. The kit is capable of identifying multiple epitopes of the cTnI antigen, so that the analytical sensitivity and precision ofthe cardiac troponin I chemiluminescence immunoassay kit are improved.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to the technical field of magnetic particle chemiluminescence immunodetection, in particular to a high-sensitivity cardiac troponin I magnetic particle chemiluminescent immunodetection kit and its preparation method and application. Background technique [0002] Troponin (troponin, Tn) complex is composed of troponin I, troponin T, and troponin C, which can regulate muscle contraction. Cardiac troponin I (cTnI) is a isoform of troponin I present in cardiomyocytes, which is highly specific for the myocardium and can be used to identify clinical situations related to myocardial injury and skeletal muscle injury. Acute myocardial infarction (AMI) is one of the major diseases threatening human life. When myocardial infarction or ischemia damages myocardial cells, cTnI quickly enters the blood through the myocardial cell membrane. Therefore, rapid, sensitive and accurate dete...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/531
Inventor 陈小茹肖桥斌吴向东周博
Owner SHENZHEN AMTECH BIOENGINEERING LTD INC
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