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Method for detecting cardiac troponin I and application thereof

A cardiac troponin and detection kit technology, applied in the field of medical testing, can solve the problems of increased antibody dosage and production cost, loss of antibody binding force, false positive test results, etc., to improve accuracy and reliability, reduce Good dosage and stability

Active Publication Date: 2012-07-04
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, when cTnI antibody is coated on latex particles, physical adsorption method and chemical coupling method can generally be used. Physical adsorption method is less stable than chemical coupling method and is susceptible to interference from rheumatoid factor (RF) and heterophilic antibodies. Lead to false positive or falsely elevated test results
The chemical coupling method currently used is mostly random coupling method, which will lead to the loss of antibody binding force, increase the amount of antibody and production cost

Method used

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  • Method for detecting cardiac troponin I and application thereof
  • Method for detecting cardiac troponin I and application thereof
  • Method for detecting cardiac troponin I and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Selection of cTnI antibody composition

[0034] In this example, two antibody compositions were selected as the preparation of the kit of the present invention for testing. These two antibody compositions are: M18 18～28 、19C7 41～49 、560 83～93 、MF4 190～196 4 monoclonal antibodies (purchased from HyTest) were combined at a ratio of 1:1:1:1, and M18 18～28 、19C7 41～49 , 8E10 86～90 、458 169～178 Four monoclonal antibodies (purchased from HyTest) were combined at a ratio of 1:1:1:1.

[0035] Of course, in addition to the above combination of antibody compositions, other combinations of antibody compositions that meet the requirements can also be used.

Embodiment 2

[0036] The preparation of embodiment 2 cTnI detection kit

[0037] The main raw materials of this embodiment are as follows:

[0038] 1. Antibodies:

[0039] Kit 1 of the present invention: M18 18～28 、19C7 41～49 、560 83～93 、MF4 190～196 (The ratio is 1:1:1:1)

[0040] Kit 2 of the present invention: M18 18～28 、19C7 41～49 , 8E10 86～90 、458 169～178 4 (the ratio is 1:1:1:1)

[0041] Control kit: M18 18～28 、19C7 41～49 (The ratio is 1:1)

[0042] 2. Latex:

[0043] The present invention is only exemplarily using polystyrene latex particles with a diameter of 150-250 nm to carry out experiments with carboxyl groups.

[0044] The main reagents of this embodiment are prepared as follows:

[0045] Reagent R1: Glycine buffer solution containing 1.2% PEG6000 (polyethylene glycol 6000), 0.15M (mol / L) NaCl. The reagent is a colorless transparent solution.

[0046] Reagent R2: Conjugate polystyrene latex particles with an anti-human cTnI antibody composition. The reagent is...

Embodiment 3

[0058] Example 3 Determination of cTnI

[0059] Detection tool: Hitachi 7060 automatic biochemical analyzer.

[0060] Analysis method: two-point endpoint method; wavelength: 600nm; sample volume: 25ul; R1: 200ul; R2: 50ul; calibration method: multi-point calibration; reaction direction: upward; measurement temperature: 37°C; Read the absorbance A at the first minute 1 ;Add R2 at 5min, read absorbance A at 4min 2 . The reaction absorbance was calculated as the calibrated difference between A2 and A1.

[0061] Calculation method: multi-point calibration, multi-parameter curve equation (such as logit / log) fitting, and ΔA can be used to obtain troponin content.

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Abstract

The invention provides a method for detecting cardiac troponin I (cTnI) and a cardiac troponin I detection kit prepared by applying the method. The method comprises the steps of coupling an antibody composition with polystyrene latex particles, then subjecting the coupled material to immunoreaction with corresponding antigen in a sample to be detected to form aggregated particles, and determining turbidity generated by the reactant under a wavelength of 400-700nm to obtain the content of cTnI in the sample to be detected. The antibody composition consists of at least three antibodies, and the antibodies are specific to cTnI without cross reaction with fast skeletal muscle troponin I and slow skeletal muscle troponin I. In addition, if one antibody in the antibody composition is sensitive to a certain factor influencing the accurate determination of cTnI in the sample to be detected, and then at least another antibody is not sensitive to the factor influencing the accurate determination of cTnI. The method of the invention is simple to operate, and has a wide scope of application, high precision, strong capability of resisting disturbance and low production cost.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a method for detecting cardiac troponin I and a cardiac troponin I detection kit prepared by using the method. Background technique [0002] Currently, the main biochemical markers for detection of acute myocardial infarction (AMI) are creatine phosphokinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), cardiac troponin T (cTnT) and cardiac troponin I ( cTnI) and so on. CK-MB and LDH are widely distributed in the body, so their specificity is low, and many diseases can cause their elevation, and their maintenance time in the blood is short; cTnT has a lot of homology with skeletal muscle troponin T (sTnT) High, prone to cross-reaction. cTnI has the advantages of high specificity, early appearance, high sensitivity, and long duration in the blood. It can be used as an early diagnostic index for AMI and has been proven to be one of the most specific and sensitive serum ...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 邹炳德邹继华夏佳音
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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