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Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum

A low-density lipoprotein and quantitative determination technology, which is applied in the field of quantitative determination reagents and determination kits for serum low-density lipoprotein cholesterol, can solve the problems of clinical misleading, no substantial improvement, and no popularization and use, and achieves high sensitivity , low cost effect

Inactive Publication Date: 2012-07-04
宁波天康生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the TC, TG and HDL-C contents of a serum sample are all within the normal range, and the determination of the above three lipids has good accuracy, the LDL-C content calculated according to the Friedewald formula is generally also It can be considered accurate; however, the Friedewald formula calculation method cannot be applied to serum samples of patients with dyslipidemia, especially hypertriglyceridemia, and the subjects who need to determine LDL-C in clinical practice often have dyslipidemia. The LDL-C content of the patient's serum sample is calculated by the Friedewald formula calculation method, and the obtained results are meaningless for clinical diagnosis and treatment, and are likely to cause misleading clinical results
[0006] In the early 1990s, the second generation of precipitation and separation of LDL based on the principle of antigen-antibody immunoassay briefly appeared. It only replaced the centrifugation step with a new method on the precipitation method and even on the sample, which was not substantive. Improvement of this kind of method has not been promoted and used in China

Method used

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  • Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum
  • Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum
  • Reagent and kit for quantitatively determining low-density lipoprotein cholesterol (LDL-C) in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] One, prepare following reagent 1 and reagent 2 of the present invention according to following composition and ratio:

[0043] Reagent 1:

[0044]

[0045] Reagent 2:

[0046]

[0047]

[0048] 2. The determination uses the HITACHI 7100 automatic biochemical analyzer, and the analysis parameters are set as follows: analysis method: endpoint method; measurement point: 16, 34; detection wavelength: 570nm (main) / 700nm (secondary); reaction temperature: 37°C; Sample volume: 3.0 μl; reagent R1: 210 μl; reagent R2: 70 μl; reaction direction: ascending; calibration mode: linear mode, 2-point calibration; unit: mmol / L. The instrument automatically performs the following operation process: first add 3 μl of the serum sample to be tested into the cuvette, then add 210 μl of reagent 1, mix well, keep it in a constant temperature environment at 37°C for 5 minutes, and read the absorbance A1; then add the reagent 2 Start the second enzyme reaction, continue to incubate, a...

Embodiment 2

[0059] Using the reagents listed in Example 1, the LDL-C content of 20 serum samples was measured according to the method and conditions described in Example 1, and each serum sample was simultaneously tested with a commercially available LDL-C assay kit (Shanghai The product of Zixing Pharmaceutical Company, batch number M081209) carries out comparative timing determination, and the results are listed in Table 2 below:

[0060] Table 2 The assay result of reagent of the present invention and contrast reagent to LDL-C

[0061]

[0062] After statistical analysis of the experimental data, we get:

[0063] Related equation: Y=1.0099x+0.0434 (n=20)

[0064] R 2 =0.9961 or R=0.9980 (analyzed by statistical analysis software)

[0065] The accuracy of the method of the present invention is measured by a correlation experiment with the control method. Gained correlation equation Y=1.0099x+0.0434 from the statistical result of experimental data can find out that the slope is cl...

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Abstract

The invention provides a reagent for directly and quantitatively determining low-density lipoprotein cholesterol (LDL-C) in human serum in a homogeneous system by an enzymatic method. The reagent is suitable for automatically and quantitatively determining the LDL-C by an automatic biochemical analyzer. The reagent consists of a solution type reagent 1 and a solution type reagent 2 which are placed separately, wherein the reagent 1 contains cholesterol esterase, cholesterol oxidase, 4-amino-antipyrine, catalase, surfactant, a buffering agent and a stabilizing agent; and the reagent 2 comprises peroxidase, a color-developing agent, the other surfactant, a stabilizing agent and a buffering agent. The invention also provides a method for directly determining the content of the LEDL-C in the homogeneous system without pre-treating a serum sample, and a kit which specifically uses the method in a clinical laboratory and in which the reagent 1 and the reagent 2 are accommodated.

Description

technical field [0001] The invention relates to a reagent and kit for quantitative determination of low-density lipoprotein cholesterol (LDL-C) in serum samples. Background technique [0002] Lipids in serum such as cholesterol and triglycerides do not exist in the serum in free form, but exist in the serum in the form of lipoproteins after being combined with various apolipoproteins. Lipoproteins in serum are particles According to particle size and density, it can be divided into four types of lipoproteins: chylomicrons (CM), very low-density lipoprotein (VLDL), low-density lipoprotein (LDL) and high-density lipoprotein (HDL). Lipoproteins play different physiological and pathological functions in the body. Since the 1970s, numerous epidemiological studies have confirmed that serum low-density lipoprotein cholesterol (LDL-C) levels are associated with atherosclerosis or coronary heart disease. The incidence is positively correlated, and it has been recognized that low-den...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N21/78
Inventor 李清华宋高峰
Owner 宁波天康生物科技有限公司
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