In-vitro diagnostic reagent for homogeneous method of low-density lipoprotein cholesterol (LDL-C) of serum

Abstract

The invention relates to an in-vitro diagnostic reagent for a homogeneous method of low-density lipoprotein cholesterol (LDL-C) of serum, wherein the in-vitro diagnostic reagent is capable of being widely applied to the technical field of medicine and biochemistry and is characterized in that the in-vitro diagnosis is carried out by means of a method comprising the following steps of: step one, selectively cracking chyle particles (CM), very low density lipoprotein cholesterol (VLDL-C) and high density lipoprotein cholesterol (HDL-C) within the serum by using a group of surfactant comprising trimethyl-beta-cyclodextrin, ethylene oxide octadecyl amine, poloxamer F88 and Brij-58, then generating hydrogen peroxide (H2O2) during the catalytic reaction of cholesterol esterase (COE) and cholesterol oxidase (COD), and then discomposing the H2O2 by means of a chemiluminescence clearing system of hydrogen peroxide, wherein the LDL-C particles within the serum are still kept perfectly at the moment; step two, reacting the LDL-C by catalyzing with the COE and the COD under the effect of TritonX-100 so as to generate H202, then promoting a chemiluminescence quantitative system to produce chemiluminescence by catalyzing the H2O2 with POD, and quantitating the LDL-C after measuring luminous intensity. The measuring reagent provided by the invention has the advantages that the sensitivity is high, the capacity of resisting disturbance is strong, the purpose for detecting the LDL-C of serum in batch is realized on a microporous plate chemiluminescence apparatus by measuring chemiluminescence intensity, and the reagent is suitable for the application in clinical laboratory.

Description

technical field [0001] The invention relates to an in vitro diagnostic reagent for serum low-density lipoprotein cholesterol (LDL-C) homogeneous method, which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] LDL-C is a kind of lipoprotein with the most cholesterol content in plasma, the particles are small, and its cholesterol content (including cholesteryl ester and free cholesterol) is more than half. The main physiological function of LDL-C is to transport the cholesterol in the liver to various tissues for utilization through the blood, and promote the massive accumulation of cholesterol in the cells. Numerous epidemiological studies have shown that serum LDL-C is positively correlated with atherosclerosis (AS) and coronary heart disease (CHD). [0003] The clinical methods for measuring LDL-C include ultracentrifugation precipitation method, Friedewald formula method, various electrophoresis methods, and the referen...

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Problems solved by technology

The disadvantage of these methods is that the detection depends on the automatic biochemical analyzer, not only the test sensitivity is low, but also in order to meet the requirements of the parameter setting of the automatic biochemical analyzer, even if the minimum sample volume allowed by the instrument is used, there are still problems for the determination system. Excessive amount of specimens taken, too many interfering substances coexisting with LDL-C, in addition, the complex formed by phosphotungstate, or a single surfactant, or a combination of phosphotungstate and a single surfactant and LDL-C There is a tendency to decompose during the process of clearing serum non-LDL-C, so the interference of non-LDL-C components in serum cannot be completely ruled out

[0005] Chemiluminescence analysis is an analytical method based on the phenomenon of chemiluminescence. This method does not require complex instruments, light sources (thus reducing or eliminating Rayleigh scattering and Raman scattering) and dispersion devices, and there is no visible light colorimetry. The interference of scattered light and stray light, which is common in quantitative analysis methods, greatly improves the signal-to-noise ratio, so it has the advantages of high sensitivity and wide linear range. It has been widely used in life sciences, clinical medicine, environmental testing, and food analysis in recent years. and various fields such as agricultural analysis, but has not been applied to the quantitative analysis of serum LDL-C

The LDL-C quantitative analysis method applied to the automatic biochemical analyzer cannot be applied to LDL-C due to the difference in the buffer capacity and pH value of the buffer system, and the determination reagent contains substances that inhibit chemiluminescence such as catalase and sodium azide. Establishment of Chemiluminescent Quantitative Detection Reagents

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