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Small and dense low-density lipoprotein cholesterin detection kit and preparation thereof

A low-density lipoprotein and detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of inaccurate determination of concentration, poor specificity, and inaccurate determination of chemical precipitation method.

Inactive Publication Date: 2015-06-03
王贤俊
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gradient density ultracentrifugation, molecular sieve chromatography, capillary electrophoresis, and dynamic light scattering are cumbersome to operate, and have high requirements for laboratory conditions and experimental operators. Gradient gel electrophoresis is time-consuming and has poor specificity. The measured value of chemical precipitation method Inaccurate, nuclear magnetic resonance spectroscopy needs to be equipped with expensive experimental equipment, index evaluation method can only be roughly estimated, and its concentration cannot be accurately determined

Method used

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  • Small and dense low-density lipoprotein cholesterin detection kit and preparation thereof
  • Small and dense low-density lipoprotein cholesterin detection kit and preparation thereof
  • Small and dense low-density lipoprotein cholesterin detection kit and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1 The preparation of small and dense low-density lipoprotein cholesterol (sdLDL-C) detection kit (1)

[0027] Concrete composition is as follows (drugs and enzymes used in the following reagents are all commercially available):

[0028]

[0029] 1) Determination of precision: 20 consecutive samples were taken for measurement, and the mean, standard deviation and coefficient of variation of the measured values ​​were calculated.

[0030]

[0031] Table 1 Precision test results

[0032]

[0033]

[0034] The coefficient of variation CV is usually used to measure the precision of a determination method, the smaller the CV value, the better the precision of the result of the determination method. For clinical chemistry tests, a method precision with a CV of less than 5% is generally accepted as acceptable. The CV value in Table 1 is less than 3%, indicating that the method of the present invention has excellent precision.

[0035] 2) Measurement of...

Embodiment 2

[0043] Embodiment 2 The preparation of small and dense low-density lipoprotein cholesterol (sdLDL-C) detection kit (2)

[0044] Concrete composition is as follows (drugs and enzymes used in the following reagents are all commercially available):

[0045]

[0046] The results show that the precision, accuracy, linear correlation and analytical sensitivity are within the normal error range (data omitted) compared with the data of the reagent described in Example 1.

Embodiment 3

[0047] Embodiment 3 The linear correlation determination of reagent of the present invention and commercially available reagent A:

[0048] Concrete composition is as follows (drugs and enzymes used in the following reagents are all commercially available):

[0049]

[0050]

[0051] Respectively adopt reagent of the present invention and commercially available reagent A, adopt Olympus 400 full-automatic biochemical analyzer, to 50 samples (comprising normal and abnormal sample), measure by respective parameter, and carry out correlation analysis to measured value (result See figure 2 , the X-axis represents the measured value of the test of the present invention, and the Y-axis represents the measured value of the commercially available reagent A). Correlation coefficient: r 2 =0.9930, the linear equation is: y=0.99x+0.008, the results show that this reagent has a good correlation with commercially available reagents.

[0052] Table 4 Linear correlation detection resu...

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PUM

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Abstract

The invention provides a small and dense low-density lipoprotein cholesterin (sdLDL-C) detection kit which comprises double liquid reagents, including a reagent R1 and a reagent R2, wherein the reagent R1 comprises an MOPS buffer liquid (the pH value is 7.0), cholesterol esterase, cholesterol oxidase, phospholipase, catalase, polyoxyethylene alkyl phenyl ether (JK-14) as a surfactant A, polyoxyethylene benzyl styrene ether (A3PK) as a protecting agent, and N-ethyl-N-(2-hydroxyl-3-sulfopropyl)-3-methylaniline sodium salt (TOOS); the reagent R2 comprises an MOPS buffer liquid (the pH value is 7.0), peroxidase, polyethylene glycol octylphenol ether (Triton X-100) as a surfactant, 4-amino antipyrine (4-AA), and sodium azide. The method provided by the invention is high in sensitivity, high in specificity, simple in reagent preparation, free of sample pretreatment, and worthy of popularization and application.

Description

Technical field: [0001] The invention provides a small and dense low-density lipoprotein cholesterol (sdLDL-C) detection kit, the kit is a liquid double reagent, including reagents R1 and R2, the reagent R1 contains MOPS buffer (PH7.0) 20~80mmol / L, cholesterol esterase 1400~1800U / L, cholesterol oxidase 2300~2600U / L, phospholipase 2500~3000U / L, catalase 1000~1300U / L, polyethylene as surfactant A Glycol alkyl phenyl ether (JK-14) 1.0~3.0g / L, polyoxyethylene benzyl styrene ether (A3PK) as a protective agent 0.1~0.5g / L, N-ethyl N-(2- Hydroxy-3-sulfopropyl)-3-methylaniline sodium salt (TOOS) 0.3~0.7mmol / L; Reagent R2 contains MOPS buffer (PH7.0) 20~80mmol / L, peroxidase 2000~5000U / L, polyethylene glycol octyl phenyl ether (Triton X-100) 5-18g / L as surfactant B, 4-aminoantipyrine (4-AA) 25-60mmol / L, azide Sodium 0.05%. This reagent has high sensitivity, strong specificity, simple reagent preparation, and no pretreatment of samples, so it is worthy of further promotion and use. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/60C12Q1/44C12Q1/30C12Q1/28C12Q1/26
Inventor 王贤俊
Owner 王贤俊
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