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Homogeneous detection kit for creatine kinase isoenzyme and application of thereof

A detection kit, creatine kinase technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of low sensitivity, inability to take into account high sensitivity and wide detection range, etc., and achieve suitable sensitivity and wide detection range. Effect

Pending Publication Date: 2021-07-16
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] CN106093411A discloses a one-step homogeneous CK-MB detection kit for the diagnosis of acute myocardial infarction and its application in detecting CK-MB content. Fast, strong specificity and other advantages, but the sensitivity is still not high, and this kit can not take into account high sensitivity and wide detection range

Method used

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  • Homogeneous detection kit for creatine kinase isoenzyme and application of thereof
  • Homogeneous detection kit for creatine kinase isoenzyme and application of thereof
  • Homogeneous detection kit for creatine kinase isoenzyme and application of thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] The preparation of embodiment 1 reagent 1

[0092] 1.1 Preparation and characterization process of the first carrier

[0093] 1) Prepare a 100ml three-neck flask, add 40mmol styrene, 5mmol methacrylic acid, 10ml water, stir for 10min and pass N 2 30min;

[0094] 2) Weigh 0.12g of potassium persulfate and 0.2g of sodium chloride, dissolve them in 40ml of water to prepare an aqueous solution. This aqueous solution is added in the reaction system of step 1, continues to pass through N 2 30min;

[0095] 3) The temperature of the reaction system was raised to 70° C., and the reaction was carried out for 15 hours;

[0096] 4) Cool the emulsion after the reaction to room temperature, and filter with a suitable filter cloth. The obtained emulsion is washed with deionized water for several times by centrifugal sedimentation until the conductivity of the supernatant at the beginning of centrifugation is close to that of deionized water, then diluted with water, and preserv...

Embodiment 2

[0120] The preparation of embodiment 2 reagent 2

[0121] 2.1 Dialyze CK-MB Antibody II to 0.1M NaHCO 3 Buffer, the measured concentration is 1.1mg / ml.

[0122] 2.2 Take 0.5 mg of CK-MB antibody II in step 1), adjust the concentration to 1 mg / ml, add 2.7 μl of biotin with a concentration of 16.8 mg / ml, mix well, and react at 2-8°C for 14-18 hours.

[0123] 2.3 After the reaction is completed, dialyze in 20 mM Tris buffer.

Embodiment 3

[0124] The preparation of embodiment 3 reagent 3

[0125] 3.1 Preparation of the second carrier

[0126] a) Prepare a 100ml three-neck flask, add 40mmol styrene, 5mmol acrolein, and 10ml water, stir for 10min, and pass N 2 30min.

[0127] b) Weigh 0.11g of ammonium persulfate and 0.2g of sodium chloride and dissolve them in 40ml of water to prepare an aqueous solution. This aqueous solution is joined in the reaction system of step a), continues to pass through N 2 30min.

[0128] c) The temperature of the reaction system was raised to 70° C. and reacted for 15 hours.

[0129] d) Cool the emulsion after the reaction to room temperature, and filter with a suitable filter cloth. The obtained emulsion is washed with deionized water for several times by centrifugation and sedimentation until the conductivity of the supernatant at the beginning of centrifugation is close to that of deionized water, then diluted with water, and stored in the form of emulsion.

[0130] e) The ...

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Abstract

The invention relates to a homogeneous detection kit for creatine kinase isoenzyme and application thereof. The kit comprises a reagent 1 and a reagent 3, wherein the reagent 1 comprises a first buffer solution and acceptor particles suspended in the first buffer solution, the reagent 3 comprises a second buffer solution and donor particles suspended in the second buffer solution, the acceptor particles comprise a first carrier, the first carrier is filled with a luminous composition, and a particle size distribution variable coefficient C.V value of the receptor particles in the reagent 1 is not lower than 5% and not higher than 25%; and the donor particles comprises second carriers, the second carriers are filled with a sensitizing agent, and the sugar content of the donor particle in each milligram of the donor particles in the reagent 3 is not higher than 40 mg. The homogeneous detection kit provided by the invention can be used for clinical auxiliary diagnosis of myocardial infarction, and not only has relatively proper sensitivity, but further has a very wide detection range.

Description

technical field [0001] The invention belongs to the technical field of homogeneous detection, and in particular relates to a homogeneous detection kit of creatine kinase isoenzymes, a preparation method and application thereof. Background technique [0002] Creatine kinase (CK) is a dimeric enzyme that has four different forms: mitochondrial isozyme, cytoplasmic isozyme CK-MM (muscle type), creatine kinase brain isozyme CK- BB (brain type), creatine kinase isoenzyme (CK-MB), in which creatine kinase isoenzyme is mainly located in the myocardium. [0003] Creatine kinase isoenzyme is released into the peripheral blood during acute myocardial infarction (AMI) and myocardial necrosis, and can be detected 3-8 hours after the onset of clinical symptoms. Combined detection with myoglobin and troponin can judge myocardial Whether it is damaged is an index for researching and diagnosing acute myocardial infarction (AMI). Clinically, it is mainly used for auxiliary diagnosis of myo...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/573G01N33/577
CPCG01N33/54313G01N33/573G01N33/577
Inventor 许家琦杨阳李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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