Blood sample assay method
First Claim
1. An enzymatic method not requiring a high density lipoprotein (HDL) calibration standard for measuring the concentration of cholesterol, associated with a high density lipoprotein (HDL) component in a plasma portion of a blood-cell containing sample, wherein the sample contains both a HDL component and a non-HDL component, which also comprises cholesterol, wherein the HDL component is a component devoid of apolipoprotein B, and wherein the non-HDL component is a component containing apolipoprotein B in the plasma portion of the blood-cell containing sample, the method comprising the steps of:
- i) diluting the sample;
ii) substantially removing blood cells to provide a substantially cell free sample;
iii) contacting the sample with a blocking reagent for the non-HDL component such that access to cholesterol from the non-HDL component is temporarily blocked while cholesterol from the HDL component remains accessible to a cholesterol converting enzyme;
iv) contacting the sample with at least one converting enzyme that reacts specifically with the cholesterol associated with the HDL and generates one or more detectable reaction product(s);
v) monitoring the detectable reaction product(s);
vi) relating an amount of the detectable product(s) to the concentration of the cholesterol associated with the HDL component in the blood sample, wherein the concentration of the cholesterol associated with the HDL component is related to the amount of the corresponding detectable reaction product(s) by means of estimating an un-measurable (fictive) endpoint for the concentration of the HDL-associated cholesterol by an algorithm;
wherein step iii) may be carried out at any stage up to and including step iv) but before steps v) or vi) and wherein the blocking reagent of step iii) may be added to the sample separately or in step i) or step iv), and wherein steps i) and iii) are carried out before step ii).
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Abstract
The invention provides an enzymatic method for measuring the concentration of one or more analytes in the plasma portion of a blood derived sample, containing a first and a second component, where said second component interferes with the measurement of said first component. The method includes: i) diluting the sample with a reagent mixture; ii) substantially removing blood cells; iii) using a reagent which serves to temporarily prevent reaction of the second component, to generate a blocked second component; iv) causing the selective reaction of a constituent of each analyte to directly or indirectly generate detectable reaction products, where one of the analytes is the first component; v) monitoring the detectable reaction product or products; vi) relating an amount of the detectable product or products and/or a rate of formation of the detectable product or products to the concentration of each analyte, where the concentration of at least the first component is related to a corresponding detectable reaction product by means of estimating an un-measurable (fictive) endpoint. Step iii) may be carried out at any stage up to and including step iv) but before steps v) or vi). The reagent of step iii) may be applied to the sample separately or may be included in a reagent mixture during steps i) or iv). A corresponding kit is also provided.
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Citations
21 Claims
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1. An enzymatic method not requiring a high density lipoprotein (HDL) calibration standard for measuring the concentration of cholesterol, associated with a high density lipoprotein (HDL) component in a plasma portion of a blood-cell containing sample, wherein the sample contains both a HDL component and a non-HDL component, which also comprises cholesterol, wherein the HDL component is a component devoid of apolipoprotein B, and wherein the non-HDL component is a component containing apolipoprotein B in the plasma portion of the blood-cell containing sample, the method comprising the steps of:
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i) diluting the sample; ii) substantially removing blood cells to provide a substantially cell free sample; iii) contacting the sample with a blocking reagent for the non-HDL component such that access to cholesterol from the non-HDL component is temporarily blocked while cholesterol from the HDL component remains accessible to a cholesterol converting enzyme; iv) contacting the sample with at least one converting enzyme that reacts specifically with the cholesterol associated with the HDL and generates one or more detectable reaction product(s); v) monitoring the detectable reaction product(s); vi) relating an amount of the detectable product(s) to the concentration of the cholesterol associated with the HDL component in the blood sample, wherein the concentration of the cholesterol associated with the HDL component is related to the amount of the corresponding detectable reaction product(s) by means of estimating an un-measurable (fictive) endpoint for the concentration of the HDL-associated cholesterol by an algorithm; wherein step iii) may be carried out at any stage up to and including step iv) but before steps v) or vi) and wherein the blocking reagent of step iii) may be added to the sample separately or in step i) or step iv), and wherein steps i) and iii) are carried out before step ii). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21)
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Specification
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- Resources
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Current AssigneeAbbott Rapid Diagnostics International Unlimited Company (Abbott Laboratories)
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Original AssigneeAxis-Shield PLC (Abbott Laboratories)
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InventorsFaaren, Arne Ludvig, Nordhei, Arne Kristian, Frantzen, Frank, rning, Lars, Sundrehagen, Erling
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Primary Examiner(s)Kosson, Rosanne
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Application NumberUS14/357,561Publication NumberTime in Patent Office1,698 DaysField of SearchUS Class CurrentCPC Class CodesC12Q 1/60 involving cholesterolC12Q 1/61 involving triglyceridesG01N 33/92 involving lipids, e.g. chol...
