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Chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum/plasma and preparation method thereof

A creatine kinase, quantitative detection technology, applied in the field of immunoassay, can solve the problems of low sensitivity, long reaction time, expensive reagents and instruments, etc., and achieve the effects of improving detection sensitivity, reducing reaction time, and reducing reagent cost

Inactive Publication Date: 2019-03-19
DIRUI MEDICAL TECH CO LTD
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The above method for quantitative detection of creatine kinase isoenzyme has the following defects: 1) ELISA method adopts microwell plate as immobilization, and coating density is limited, and sensitivity is low, and reaction time is longer; Luminescence method requires high equipment requirements, expensive reagents and equipment

Method used

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  • Chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum/plasma and preparation method thereof
  • Chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum/plasma and preparation method thereof
  • Chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum/plasma and preparation method thereof

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preparation example Construction

[0051] The present invention also provides a preparation method of a chemiluminescence quantitative detection kit for detecting creatine kinase isozyme in serum / plasma, the steps are as follows:

[0052] 1) Streptavidin-coated magnetic particles

[0053]Take a certain amount of magnetic particles, wash them three times with 20mM PB buffer, then dilute to 10mg / mL, add streptavidin according to the mass ratio of magnetic particles to streptavidin 1:10-1:20, After incubating at 37°C for 18-24h, separate the magnetic beads on a magnetic separation rack, remove the supernatant, wash with 20mM PB buffer 3 times, add blocking solution (20mM PB buffer containing 2-5% BSA), 37°C After incubation for 18-24 hours, separate the magnetic beads on a magnetic separation rack, remove the supernatant, wash 3 times with 20mM PB buffer, dilute to 10mg / mL with 20mM PB buffer and save for later use to obtain streptavidin coating magnetic particles;

[0054] 2) Biotin-labeled creatine kinase isoz...

Embodiment 1

[0070] This embodiment provides a method for preparing a chemiluminescence quantitative detection kit for detecting creatine kinase isoenzymes in serum / plasma, and the specific steps are as follows:

[0071] 1) Preparation of streptavidin-coated magnetic particles

[0072] Take a certain amount of magnetic particles, wash 3 times with 20mM PB buffer, then dilute to 10mg / mL, add streptavidin according to the mass ratio of magnetic particles to streptavidin 1:15, and incubate at 37°C for 18h Afterwards, separate the magnetic beads on a magnetic separation rack, remove the supernatant, wash 3 times with 20mM PB buffer, add blocking solution (20mM PB buffer containing 2% BSA), incubate at 37°C for 18h, and place on magnetic separation Separate the magnetic beads on the rack, remove the supernatant, wash 3 times with 20mM PB buffer, dilute to 10mg / mL with 20mM PB buffer and save for later use to obtain streptavidin-coated magnetic particles;

[0073] 2) Preparation of biotin-label...

Embodiment 2

[0085] This embodiment provides a method for preparing a chemiluminescence quantitative detection kit for detecting creatine kinase isoenzymes in serum / plasma, and the specific steps are as follows:

[0086] 1) Preparation of streptavidin-coated magnetic particles

[0087] Take a certain amount of magnetic particles, wash with 20mM PB buffer three times, then dilute to 10mg / mL, add streptavidin according to the mass ratio of magnetic particles to streptavidin 1:10, and incubate at 37°C for 20h Afterwards, separate the magnetic beads on a magnetic separation rack, remove the supernatant, wash 3 times with 20mM PB buffer, add blocking solution (20mM PB buffer containing 3% BSA), incubate at 37°C for 20h, and place in magnetic separation Separate the magnetic beads on the rack, remove the supernatant, wash 3 times with 20mM PB buffer, dilute to 10mg / mL with 20mM PB buffer and save for later use to obtain streptavidin-coated magnetic particles;

[0088] 2) Preparation of biotin-l...

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Abstract

The invention belongs to the technical field of immunoassay and particularly relates to a chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum / plasma and a preparation method thereof. The chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum / plasma comprises: 1) magnetic particles coated with streptavidin; 2) biotin-labeled creatine kinase MB antibody; 3) acridinium ester-labeled creatine kinase MB antibody; 4) calibrator and quality control material. The calibrator and the quality control material contain high and low concentration points respectively. The chemiluminescent quantitative detection kit for detection of creatine kinase MB in serum / plasma provided herein employs the principle of double-antibody sandwich process as well as an avidin-biotin system and employs acridinium ester to chemically illuminate; detection sensitivity of the kit herein is greatly improved, reaction time is shortened, and agent cost is reduced.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a chemiluminescent quantitative detection kit for detecting creatine kinase isoenzyme in serum / plasma and a preparation method thereof. Background technique [0002] Creatine kinase CK is a dimer composed of two subunits, which are composed of M and B subunits encoded by different genes on chromosomes 14 and 19, respectively, and can form three isozymes, namely CKMM and CKMB and CKBB, in which the creatine kinase isoenzymes of mixed monomers are mainly derived from the myocardium (CK-MB in the myocardium accounts for 15% to 25% of the total CK). These three isozymes have the same molecular weight and catalyze the same chemical reaction, but differ in their molecular structure and origin. [0003] Creatine kinase CK is a large dimeric protein, in which the M subunit has a small N-terminal structure and a large C-terminal structure; the M-type subunit consists ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543G01N21/76
Inventor 王立英韩美玉高威孙成艳何浩会
Owner DIRUI MEDICAL TECH CO LTD
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