Method for determining cholesterol concentration in remnant lipoprotein by immunobinding direct process or immuno-precipitation separation process

A lipoprotein and cholesterol technology, applied in the field of serum determination, can solve the problems of cumbersome, difficult and time-consuming operation in ordinary laboratories

Inactive Publication Date: 2006-12-20
上海北加生化试剂有限公司 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these two methods are difficult to carry out in ordinary laboratories due to the need for special instruments, time-consuming or cumbersome operations.

Method used

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  • Method for determining cholesterol concentration in remnant lipoprotein by immunobinding direct process or immuno-precipitation separation process
  • Method for determining cholesterol concentration in remnant lipoprotein by immunobinding direct process or immuno-precipitation separation process
  • Method for determining cholesterol concentration in remnant lipoprotein by immunobinding direct process or immuno-precipitation separation process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Detection of RLP by Immunobinding Direct Method

[0045] Use reagent I and reagent II, reagent I contains: 50mmol / L Tris-HDL buffer, 60U / L cholesterol lipase, 60U / L cholesterol oxidase, 150mmol / L peroxidase, 2g / L polyethylene glycol 6000 , 11.3mL rabbit anti-human ApoB100 antiserum, 7.5mL rabbit anti-human ApoA I antiserum, reagent II contains 150mmol / L peroxidase, 0.06mmol / L 4-aminoantipyrine and 50mmol / L Tris-HDL buffer liquid. The components of reagents I and II were added to the buffer solution according to their component contents, mixed evenly, and the precipitate was filtered off, and the obtained solution was stored at 4° C. for future use.

[0046] Take out 24uL serum or plasma from the test individual, add 225uL reagent I to the serum or plasma, mix well, and place at 37°C for 5 minutes. Allow ApoB100, ApoA I antigens to fully combine with antibodies. On the 722-type spectrophotometer, use a 505nm wavelength and a 1cm optical diameter cuvett...

Embodiment 2

[0050] Example 2 Detection of RLP by Immunoprecipitation

[0051] Use 113uL rabbit anti-human ApoB100 antiserum, 71uL rabbit anti-human ApoA I antiserum and 30uL serum or plasma to mix at room temperature or 37°C for 30 minutes, centrifuge at 4000 rpm to take the supernatant, and the obtained supernatant Save for later.

[0052] Take out 24uL supernatant, add 225uL reagent I to the supernatant, mix well, and place at 37°C for 5 minutes. On the 722-type spectrophotometer, use a 505nm wavelength and a 1cm optical diameter cuvette to zero with distilled water, and measure its absorbance value A 1 .

[0053] Then, add 75uL of reagent II to it, and keep it warm at 37°C for 5 minutes. On a 722-type spectrophotometer, use a 505nm wavelength and a 1cm optical diameter cuvette to zero with distilled water, and measure its absorbance value A 2 .

[0054] Use the known concentration c.f.a.s. cholesterol calibrator as the calibration solution, take 24uL and follow the same...

Embodiment 3

[0057] Linearity, reaction process, precision, recovery rate and interference test of the detection method of the present invention

[0058] 1. Linear Analysis

[0059] Take a low-value sample (RLP-C is 0.08mmol / L) and a high-value sample (RLP-C is 2.00mmol / L) and mix them equally to prepare the RLP-C of the median sample to be 1.04mmol / L. The RLP-C of two horizontal specimens was prepared by mixing equal volumes of samples with high value and low value respectively, which were 1.52mmol / L and 0.56mmol / L respectively. Samples of 5 different levels were measured 4 times with the method of Example 1 in descending order by two clearance methods respectively, and the linear analysis was carried out according to the NCCLS EP6-P file. Results The linear regression analysis equation was: Y=0.986X+0.019, r=0.992, indicating that the method had a good linear relationship within 2.00mmol / L.

[0060] 2. Intra-batch accuracy

[0061] Take samples of 3 different levels of RLP-C...

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Abstract

The invention discloses a serum or RLP density in the plasma testing method, which is characterized by the following: adopting immune combined direct method or immune sediment separating method; providing effective estimation and reference accordance for atherosclerotic disease, coronary disease and relative metabolic disease of atherosclerotic disease.

Description

Technical field: [0001] The invention relates to a method for measuring the concentration of lipoprotein remnants (RLP) in serum or plasma, in particular to the method for measuring the concentration of lipoprotein remnants (RLP) in serum or plasma by immunobinding direct method or immunoprecipitation separation method, as a clinical judgment Atherosclerotic disease, coronary heart disease, and atherosclerotic-related metabolic disease degree and curative effect evaluation, the reference basis for prognosis judgment. Background technique: [0002] Remnant lipoprotein (RLP), also known as triglyceride (TG)-rich lipoprotein remnant (TRL), is chylomicron (chylomicron, CM) and very low density lipoprotein (very low density lipoprotein, After being hydrolyzed by lipoprotein lipase (lipoprotein lipase, LPL), triglycerides, phospholipids, apolipoprotein A (apoA, apoA) and apoC are gradually lost, and VLDL is transformed into a smaller protein rich in cholesterol, cholesteryl lipid ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/68G01N33/577
Inventor 刘颖冰朱剑锋刘颖成刘中令
Owner 上海北加生化试剂有限公司
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