Background and Overview[1][2]
Creatine phosphostearate kinase is referred to as CPK. It is mainly distributed in skeletal muscle and cardiac muscle, and also contains small amounts in brain tissue and smooth muscle. Normal human serum contains only a small amount of creatine phosphokinase. However, in certain diseases, a large amount of creatine phosphokinase may appear in the blood. For example, it is significantly elevated in children with myocarditis.
Also known as creatine kinase. CPK is an organ-specific enzyme that mainly exists in skeletal muscle, cardiac muscle and brain tissue. CPK mainly catalyzes the biochemical reaction of creatine phosphate (CP) and adenosine diphosphate (ADP) in muscles to synthesize creatine (C) and adenosine triphosphate (ATP). The serum CPK content of normal pigs is low, but due to damage to muscle cells in pigs under stress, the content and activity of CPK in the blood can increase. Therefore, CPK is often used as one of the indicators for evaluating pig stress syndrome. There are many methods for CPK determination. The commonly used method is the Antonik method. The measurement result is negative when it is below 30 units/ml, and it is positive when it is above 31 units/ml. In addition, there is the Siqma method, which is negative when the measurement result is below 100 units/ml. , 101~200 units/ml is positive.
Measurement method[3]
Creatine kinase (CK) mainly exists in skeletal muscle and cardiac muscle, and also in brain tissue. The following conditions can increase CK activity: it begins to increase 2 to 4 hours after acute myocardial infarction, can reach 10 to 12 times the upper limit of normal, and returns to normal in 2 to 4 days. Measuring CK is better than AST, LD, etc. in diagnosing myocardial infarction; viral myocarditis is significantly increased and has reference value for its diagnosis and prognosis; patients with cerebrovascular accidents, meningitis, and hypothyroidism; strenuous exercise, various intubations, and Surgery, intramuscular injection of hibiscus and antibiotics, etc.
The commonly used method for measuring creatine kinase in serum is the colorimetric method, which uses CK to catalyze the reaction of creatine phosphate (CrP) and adenosine diphosphate (ADP) to generate adenosine triphosphate (ATP) and creatine ( Cr), hexokinase (HK) catalyzes the reaction between glucose and ATP to form glucose-6-phosphate, glucose-6-phosphate dehydrogenase catalyzes the oxidation of glucose-6-phosphate to form 6-phosphate glucose lactone and NADPH, and NADPH is generated The rate represents CK activity.
CN200610115000.1 provides a chemiluminescence determination method for creatine kinase in serum with high sensitivity, wide linear range, convenience and safety. The main technical solution adopted to solve this technical problem: the chemiluminescence determination method of creatine kinase in serum, which is characterized by using CK to catalyze the reaction of creatine phosphate (CrP) and adenosine diphosphate (ADP) to generate adenosine triphosphate (ATP) and creatine (Cr); then glycerol kinase (GK) is used to catalyze adenosine triphosphate (ATP) and glycerol to generate glycerol 3-phosphate (G-3-P); glycerol 3-phosphate (G-3-P) is oxidized by glycerol phosphate oxidase (GPO) Oxidation, and produce hydrogen peroxide; then through the action of oxidase, hydrogen peroxide oxidizes the chemiluminescent substance and emits light; the size of the light signal is positively correlated with the concentration of creatine kinase, that is, the greater the concentration of creatine kinase, the greater the light signal emitted The stronger; by recording this light signal, the concentration of creatine kinase can be inferred; using the known concentration of creatine kinase and the measured light signal, a dose-response curve is made; the creatine kinase content of the unknown sample can be estimated from this curve Come out; the chemiluminescent substance is luminol.
Compared with the existing technology, the present invention has the following advantages: it uses chemiluminescent substances instead of chromogenic substances to achieve the purpose of being more sensitive, more stable, wider and safer. This method can be used to prepare a corresponding commercial kit for quantitative determination of creatine kinase in serum PVC resin powder latest price.
The specific advantages are: ① Higher sensitivity. The traditional method is to detect the depth of color, while chemiluminescence is a photometric signal that can be counted one photon at a time. The measured minimum limit is smaller, so the sensitivity is higher. ②Wide linear range. The resolution of measuring color depth is much lower than the accuracy of photon counting, and the detection range of chemiluminescence is as high as 105. ③Safe and applicable. Chemiluminescent substances such as luminol are very safe, and no harmful substances appear during the entire detection process. It is safe whether detecting or processing waste, and is conducive to environmental protection. ④Easy to popularize.
The kit prepared by the invention can be used for fully automatic measurement, manual or semi-automatic measurement, and is suitable for use in medical and research institutions at all levels, including large, medium and small. ⑤ Huge social benefits. Chemiluminescence can be used to measure biochemical indicators or immune substances using the same chemiluminescence instrument. There is no need to import expensive biochemical instruments, which saves a lot of money. It not only reduces costs, but is of great benefit to medical institutions, patients and society.
Fixed method[2]
CN201210232877.4 proposes a method for immobilizing creatine kinase. It uses chitosan as raw material, first prepares a cross-linked chitosan solution, then prepares cross-linked chitosan microspheres, then prepares grafted lysine cross-linked chitosan resin, and finally uses grafted lysine Acid cross-linked chitosan resin was used as a carrier to immobilize creatine kinase. The invention has the advantages that conventional production equipment can be used to produce creatine phosphate under mild conditions, the process is simple, the operation is simple and easy to control, the production cost is low, the production efficiency is improved, the production process is conducive to protecting the environment; and the preparation is immobilized The stability of creatine kinase to heat, acid, alkali, organic solvents and other conditions has been significantly increased.
Main reference materials
[1] Pig Dictionary
[2] Pediatric Dictionary
[3] CN200610115000.1 Chemiluminescence determination method of creatine kinase in serum
[4] CN201210232877.4 A method of immobilizing creatine kinase
