Quantitative detection method of creatine kinase isoenzyme MB

Abstract

The invention relates to the technical field of biotechnology, in particular to a quantitative detection method of creatine kinase isoenzyme MB. The method comprises preparation of a creatine kinase isoenzyme MB solution, a microfluidic creatine kinase isoenzyme MB detection chip and a use method of the microfluidic creatine kinase isoenzyme MB detection chip. A micro-fluidic chip is structurallycharacterized by comprising a micro-pipeline network. Each working unit can independently complete single reaction or unit operation, and can also cooperate with other units to complete complex work,so that the whole chip is called a dynamic reaction system. The method has the main characteristics of high flux, miniaturization and automation, and is an innovation and leap in the aspects of traditional biotechnology, disease diagnosis and the like; immunomicrosphere chemiluminescence is combined with a micro-fluidic chip, so that a full-automatic instrument is more miniaturized and simplified,and later pipeline maintenance and mechanical arm maintenance are reduced; and the combination of the two converts the original POCT application solid-phase reaction into liquid-phase homogeneous reaction, and the method has the advantages of high reaction efficiency, high sensitivity and detection interval of the traditional tubular chemiluminiscence.

Description

technical field [0001] The invention relates to the technical field of biotechnology, in particular to a quantitative detection method for creatine kinase isoenzyme MB. Background technique [0002] Creatine kinase is a kinase that plays a vital role in intracellular energy transport and ATP metabolism. Creatine kinase isoenzyme MB (CK-MB) is an isoenzyme of creatine kinase (CK), which is produced by post-translational modification of CK and mainly exists in cardiomyocytes (and a small amount exists in bone muscle, intestinal tract, tongue, prostate and uterus), is a very important creatine kinase in the pathway of myocardial energy metabolism. CK-MB is composed of two subunits, decibels are M subunit and B subunit, M represents muscle and B represents brain, and its activity accounts for 15% to 30% of the total creatine kinase activity. In addition to CK-MB, there are two other creatine kinase isoenzymes, CK-BB in the brain and CK-MM in skeletal muscle, which have similar...

AI Technical Summary

Problems solved by technology

The quality method detects the concentration of CK-MB in the blood, and the sequence of development is radioimmunoassay CK-MB quantitative detection method, enzyme-linked immunoassay CK-MB quantitative detection method, chemiluminescence immunoassay and fluorescence Immunoassay, currently the most widely used and the best performance is the CK-MB reagent based on chemiluminescence. In addition, the CK-MB POCT kit immunoturbidimetry is also used for the detection of CK-MB mass method at present, but Its sensitivity and anti-interference are not as good as chemiluminescence

The enzyme activity method is for the detection of CK-MB enzyme activity, but its sensitivity and anti-interference are not as good as the chemiluminescence method

The enzyme activity method is to detect the enzyme activity of CK-MB. The main method is the immunosuppressive method. On the one hand, the specific anti-CK-MM antibody is used to inhibit the activity of the M subunit in CK-MB; The activity of B subunit in MB can quantitatively detect the enzyme activity of CK-MB through the reaction of B subunit in CK-MB to metabolize ATP. This method is widely used in the detection of early CK-MB due to its simple operation and quick results. , but its anti-interference ability is very poor, because the anti-M subunit antibody used in the immunosuppressive method cannot inhibit the activities of CK-BB, adenylyl cyclase and atypical CK that exist alone in the blood, which makes the result higher and cause False positives and are therefore gradually replaced by chemiluminescent reagents

In addition, in addition to the above two types of CK-MB detection methods, electrophoresis was used to detect CK-MB in the early days. This method takes a long time and has poor stability, so it has been basically abandoned at present.

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