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Small and Dense LDL Cholesterol Detection Kit
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A low-density lipoprotein and detection kit technology, applied in the biological field, can solve problems such as high requirements for operators, poor precision and accuracy, and unsuitability for clinical applications
Active Publication Date: 2020-01-07
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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Problems solved by technology
However, the steps of electrophoresis determination are cumbersome, and the precision and accuracy need to be improved, so the clinical applicability is not strong; the ultracentrifugation method is the standard method for the determination of lipoproteins, but the detection of sdLDL by this method requires an ultracentrifuge, and the detection is generally The amount is not high, and the requirements for operators are high, so this method has not been well promoted in clinical practice; although the fractional precipitation method makes the operation relatively simple, it still needs a precipitation step. The precision and accuracy of this method are relatively poor and not suitable for clinical application
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Embodiment 1
[0030] Correlation comparison
[0031] Reagents R1 and R2 were prepared according to the recipes in the table below, with a total of 6 groups.
[0032] Table 1
[0033]
[0034]
[0035] The dodecyl polypeptide of the present invention was purchased from ADEKA, a Japanese company (for details, please refer to the webpage: http: / / www.docin.com / p-1287395662.html The record on page 6 of ), the structural formula of the substance is
[0036] Among them, the sample reagent ratio, sample:reagent R1:reagent R2=3:150:50. After the sample was incubated with reagent R1 for 5 min, R2 was added, and the reaction was continued for 5 min, and then the absorbance was measured at 600 nm. As a control method, ultracentrifugation was used. For the specific method, see (Menys VC, Liu Y, Mackness M I, et al. Measurement of plasma small-denseLDL concentration by a simplified ultracentrifugation procedure and immunoassay of apolipoprotein B.[J].Clinica Chimica Acta, 2003, 334(1–2):9...
Embodiment 2
[0041] Linear range
[0042] Select a high-value sample with an sdLDL-C value of about 100 mg / dL, dilute it to different concentrations, such as 0.2, 0.4, 0.6, 0.8 times, and use the formula 3 reagent of the present invention for linear verification, and its linear range is 0 -100mg / dL, the correlation coefficient was 0.9952.
Embodiment 3
[0044] Anti-interference experiment
[0045] On the basis of formula 4, the following substances were added to its R1, 2 KU / L ascorbate oxidase, 10 g / L mannitol, 1 mM EDTA, 20 mM magnesium chloride. Check its anti-interference ability. The relevant experimental results are shown in Table 3. It can be found that the anti-interference performance of the sdLDL-C detection kit of the present invention is good, and is not affected by 5g / L hemoglobin, 30mg / dL conjugated bilirubin, and 50mg / dL vitamin C.
[0046] table 3
[0047]
[0048] .
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Abstract
The invention discloses a kit for detecting small and dense low-density lipoprotein cholesterin. The kit is characterized by comprising polyoxyethylene fatty acid ester, cholesterol esterase, cholesterol oxidase, dodecyl polypeptide and triton X-100, wherein the small and dense low-density lipoprotein cholesterin in a serum sample is protected under the action of the polyoxyethylene fatty acid ester, reaction of the small and dense low-density lipoprotein cholesterin with the cholesterol esterase and the cholesterol oxidase is inhibited to a certain extent, and the other lipoprotein cholesterols react with the cholesterol esterase and the cholesterol oxidase under the action of the dodecyl polypeptide to be eliminated; and the triton X-100 reacts with the remaining small and dense low-density lipoprotein cholesterin to detect the content thereof. The kit can be used for quickly and conveniently detecting sdLDL-C in the serum sample. The reagent formula is simple and easy to prepare and low in cost, large-scale industrial preparation can be carried out, and meanwhile, the kit is capable of efficiently, accurately and specifically detecting the small and dense low-density lipoprotein cholesterin in the sample.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a small and dense low-density lipoprotein cholesterol detection kit. 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, high-density lipoprotein cholesterol has a certain protective effect on arterial blood vessels. The higher the content, the lower the risk of cardiovascular disease, and the low-density lipoprotein cholesterol is easy to cause atherosclerosis and increase the risk of cardiovascular disease. However, recent studies have shown that...
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