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Reagent for lipid typing detection

A technology for reagents and blood lipids, used in biological testing, material testing, etc., can solve the problems of shortened pipeline, poor resolution, and narrow detection linear range, and achieve the effect of rapid response

Active Publication Date: 2018-01-09
宁波美康盛德医学检验所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the longer the pipeline, the longer the reaction time, the more fully the reaction can be carried out, but at the same time, the longer the pipeline, the worse the detection resolution, and to improve the resolution, it is necessary to shorten the pipeline, which creates a contradiction
At present, the conventional detection reagents generally need more than 120s to complete the reaction, and the detection linear range is narrow, which is not conducive to the reaction on the blood lipid typing detection instrument

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0029] Prepare the reagents according to the formula in the table below, and there are 6 groups in total.

[0030] Formulation of different reagents in table 1

[0031]

[0032] Among them, the serum sample reagent ratio, sample: reagent = 3:150. Start timing when the sample is incubated with the reagent, react for a total of 10 minutes, and monitor the change of absorbance at 546nm at the same time, at least record the absorbance at the beginning of the reaction for 60s, 120s and 600s, take the reaction end point of 600s as 100% reaction completion, and calculate the reaction at different time points Completion rate, such as reaction completion rate at 60s = absorbance at 60s / absorbance at 600s, completion rate at 120s = absorbance at 120s / absorbance at 600s. Utilize the 6 kinds of reagents prepared above for detection, and add the commercially available cholesterol reagent as a control reagent at the same time, and carry out comparative detection at the same time. The sa...

Embodiment 2

[0037] Comparison of sample test results

[0038] Since the reagent of the invention can react more quickly, it can be applied to a blood lipid typing detector.

[0039] Take 50ul of serum, add 1950ul of KBr (density adjusted to 1.21) to dilute, and mix well. Take a 5.1ml finger seal tube, add 3800ul of normal saline, and then slowly add 1200ul of the pre-diluted serum sample from the bottom. The centrifuge tube was placed in a rotor VTi-65.2 rotor and centrifuged with a Beckman ultracentrifuge. Parameters: rotating speed=65000rpm, ω2t=6.6*1010rad2 / sec, temperature=23°C, acceleration=6, deceleration=6.

[0040] The centrifuged sample is carefully moved to the VAP instrument of the present invention for detection. The detection parameters are: sample flow rate 1.0ml / min, reagent flow rate 1.0ml / min. The commercially available cholesterol reagents and the reagents of 6 formulations of the present invention were used for detection and comparison. The results are shown in Tabl...

Embodiment 3

[0045] linear experiment

[0046] Since the blood lipid typing detector can detect lipoprotein cholesterol of different components, and among them, low-density lipoprotein cholesterol accounts for the highest content of total cholesterol, so the linear range of analyzing LDL-C is used to illustrate the relationship between the reagent of the present invention and blood lipid typing The detectable linear range of the matching reagents. Get a high-value sample of LDL-C, dilute to the value of different concentrations, detect with formula of the present invention and contrast reagent, and its detection result is as shown in table 4

[0047] Table 4 Linear range

[0048]

[0049] As can be seen from Table 4, the blood lipid typing detection reagent of the present invention is obviously better than the traditional cholesterol detection reagent, and the upper limit of its linear range can reach 600mg / dL, while the common cholesterol detection reagent, in the range of 0~600mg / dL,...

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PUM

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Abstract

The invention discloses a reagent for lipid typing detection. The reagent is characterized by comprising alkylphenol polyethoxylate, Triton X-100, cholesterol esterase, cholesterol oxidase, peroxidase, phosphate buffer, phenol, 4-aminoantipyrine, and PC300. For a formula of the reagent, lipoprotein can be dissolved and release cholesterol or cholesteryl ester contained internally through additionof alkylphenol polyethoxylate and Triton X-100 in a specific ratio and in a mixing ratio, cholesterol ester can generate cholesterol under the action of cholesterol esterase, cholesterol can generatecholest-4-ene-3-ketone and hydrogen peroxide under the action of cholesterol oxidase, hydrogen peroxide acts with phenol and 4-aminoantipyrine (4-AAP) under the action of peroxidase so as to generatea colored material, and contents of different lipoproteins are calculated by checking changes of absorbance.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis, in particular to detecting the cholesterol content in lipoproteins of different densities in serum samples. Background technique [0002] Lipoproteins in blood can be divided into four categories according to their density: high-density lipoprotein (HDL), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL) and chylomicrons (CM). Different types of lipoprotein cholesterol have different abilities to cause atherosclerosis. Usually the most clinically detected components are high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C). Conventionally speaking, high-density lipoprotein cholesterol has a certain protective effect on arteries. The higher the content, the lower the risk of cardiovascular disease, and the low-density lipoprotein cholesterol can easily cause atherosclerosis and increase the risk of cardiovascular disease. However, recent s...

Claims

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Application Information

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IPC IPC(8): G01N33/92
Inventor 邹炳德邹继华汪屹贾江花
Owner 宁波美康盛德医学检验所有限公司
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