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Small and dense low-density lipoprotein cholesterol detection test kit and detection method thereof

A low-density lipoprotein and detection kit technology, applied in the field of biochemistry, can solve the problems of high results, too much enzyme can not be added, false positives, etc., to improve stability and avoid false positives.

Active Publication Date: 2020-09-11
NINGBO RUI BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the protective agents in domestic sd LDL-C detection kits are basically non-ionic surfactants. The principle of these non-ionic surfactants to protect small and dense low-density lipoprotein cholesterol is to reduce the reactivity of enzymes to it. But it cannot be completely inhibited. In the above situation, the corresponding enzyme should not be added too much. If there is too much, the small and dense LDL cholesterol in the low-value sample may be consumed, resulting in low results. If the amount of enzyme Insufficient, generally normal samples have no effect, but when testing samples with high values ​​of ordinary low-density lipoprotein cholesterol, non-small and dense low-density lipoprotein cholesterol cannot be completely consumed, which will lead to high results
Similarly, there are various stabilizers for the determination of low-density lipoprotein cholesterol at present, and the stability effects are different, and there may be false positives due to unstable reagents

Method used

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  • Small and dense low-density lipoprotein cholesterol detection test kit and detection method thereof
  • Small and dense low-density lipoprotein cholesterol detection test kit and detection method thereof
  • Small and dense low-density lipoprotein cholesterol detection test kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] (1) Preparation of reagents:

[0050] Preparation reagent 1: 1% (mass percentage) cyclodextrin, 1000U / L cholesterol lipase (CHER), 500U / L cholesterol oxidase (CHOD), 50U / L phospholipase, 1% (volume percentage) stabilizer Prionex, 0.5ml / L preservative ProClin300, 5kU / L peroxidase, 5mM / L chromogen TOOS and GOOD's buffer HEPES 50mM / L.

[0051] Preparation reagent 2: including 5mM / L 4AAP, Triton X-405 1% (volume percentage) and GOOD's buffer HEPES 50mM / L.

[0052] (2) Detection method

[0053] Using this kit for detection, the detection instrument can obtain different machine parameters according to different detection instruments. The reaction temperature is 37°C, the sample volume is 3 μl, the volume of reagent 1 is 150 μl, and the reaction time is 5 minutes. The absorbance of the sample is measured, and then reagent 2 is added. The volume is 50 μl, and the detection time is 5 minutes at a constant temperature of 37°C. The absorbance of the sample is measured. The main / ...

Embodiment 2

[0055]A kind of raw material of small and dense low-density lipoprotein cholesterol detection kit, including: Reagent 1: 50mM / L HEPES buffer solution, 1% (mass percentage) d-cyclodextrin, 1000U / L cholesterol lipase, 500U / L cholesterol Oxidase, 50U / L phospholipase, 1% (volume percentage) Prionex, 0.5ml / L ProClin300, 5kU / L peroxidase and 5mM / L TOOS, reagent 2: 50mM / L HEPES, 1% (volume percentage) Triton X-405 and 5mM / L 4AAP.

[0056] Use this kit for detection, the detection instrument is Hitachi automatic biochemical analyzer 7180, the reaction temperature is 37°C, the sample volume is 3 μl, the volume of reagent 1 is 150 μl, the reaction time is 5 minutes, the absorbance of the sample is measured, and then the volume of reagent 2 is 50 μl , 37 ℃ constant temperature detection time 5min, measure the absorbance of the sample, the main / secondary wavelength selected in the above absorbance measurement is 600 / 700nm, the measurement method is the two-point end point method, and the ...

Embodiment 3

[0059] A kind of raw material of small and dense low-density lipoprotein cholesterol detection kit, including: Reagent 1: 50mM / L HEPES, 1% (mass percentage) β-cyclodextrin, 1000U / L cholesterol lipase, 500U / L cholesterol oxidase , 50U / L phospholipase, 1% (volume percentage) Prionex, 0.5ml / L ProClin300, 5kU / L peroxidase and 5mM / L TOOS, reagent 2: 50mM / LHEPES, 1% (volume percentage) Triton X-405 and 5mM / L 4AAP.

[0060] Use this kit for detection, the detection instrument is Hitachi automatic biochemical analyzer 7180, the reaction temperature is 37°C, the sample volume is 3 μl, the volume of reagent 1 is 150 μl, the reaction time is 5 minutes, the absorbance of the sample is measured, and then the volume of reagent 2 is 50 μl , the reaction temperature is 37°C, the detection time is 5min, and the absorbance of the sample is measured. The main / secondary wavelengths selected when measuring the absorbance above are 600 / 700nm, the measurement method is the two-point end point method...

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Abstract

The invention discloses a small and dense low-density lipoprotein cholesterol detection test kit and a detection method thereof. The test kit comprises a reagent 1 and a reagent 2, wherein the reagent1 contains a GOOD's buffer solution, a protective agent cyclodextrin, cholesteryl esterase, cholesterol oxidase, phospholipase, peroxidase, a stabilizer prionex, a preservative pc300 and a chromogenTOOS; the reagent 2 is prepared from a GOOD's buffer solution, triton X-405 and 4AAP. The detection method of the test kit comprises the following steps: treating a reagent 1 and a sample for a certain period of time under a specific condition according to a certain ratio, then adding a reagent 2 according to a certain ratio, treating for a certain period of time under the specific condition, andquantitatively determining the content of small and dense low-density lipoprotein cholesterol according to the change of an absorbance value. The small and dense low-density lipoprotein cholesterol detection test kit disclosed by the invention contains a cyclodextrin protective agent and a special stabilizer, is particularly high in detection accuracy and good in stability for high-value samples with relatively high concentrations of low-density lipoprotein cholesterol and small and dense low-density lipoprotein cholesterol, and can meet clinical examination requirements.

Description

technical field [0001] The invention belongs to the technical field of biochemistry, in particular to a small and dense low-density lipoprotein cholesterol detection kit and a detection method thereof. Background technique [0002] Clinically, abnormal fluctuations in blood lipids often lead to the occurrence of coronary heart disease (CAD). Low-density lipoprotein (low-density lipoprotein, LDL) is recognized as a representative of lipoproteins that cause atherosclerosis; major blood lipid management guidelines at home and abroad all emphasize that LDL cholesterol (LDL cholesterol, LDL-C) is the key to reducing the risk of CAD. Key intervention targets. Recent studies have shown that among LDL subtypes, small dense LDL (sd LDL) with high density and small particles is more closely related to atherosclerosis; sd LDL or sd LDL cholesterol (sd LDL cholesterol, sd LDL-C) level is more sensitive than LDL or LDL-C as an index to assess and predict CAD risk. [0003] At present,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/60C12Q1/44C12Q1/34C12Q1/28C12Q1/26
CPCC12Q1/60C12Q1/28C12Q1/44C12Q1/26C12Q1/34G01N2333/908G01N2333/916G01N2333/902
Inventor 谢蒙张闻陈媛周海滨
Owner NINGBO RUI BIO TECH
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