US 2014/0170688 A1 · RAPID, LOW-SAMPLE-VOLUME CHOLESTEROL AND TRIGLYCERIDE ASSAYS

Abstract

Reagents, assays, methods, kits, devices, and systems for rapid measurement of cholesterol and cholesterol sub-fractions from a blood sample are provided. Total cholesterol, low density lipoprotein cholesterol, and high density lipoprotein cholesterol can be measured in a single assay using kinetic measurements, under conditions in which cholesterol sub-species are converted to a detectable product at distinct rates. The detectable product is measured at different times after assay initiation. A lipase, cholesterol esterase, cholesterol oxidase and a peroxidase may be used together to produce colored product in amounts directly proportional to the quantity of cholesterol converted. Methods for calculating very-low density lipoprotein cholesterol levels by further including triglyceride measurements are disclosed. Assays may be performed in a single reaction mixture, allowing more accurate and precise cholesterol determinations, including ratios of cholesterol sub-fractions to total cholesterol, at less expense, than would be expected by performing several different assays in different reaction mixtures.

Claims27

Dependent claims

A method for the measurement of at least two lipoprotein components in a blood sample from a subject, wherein said at least two lipoprotein components are selected from total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C), said method comprising:

combining at least a portion of said sample of blood with a first reagent to provide a first combined solution, wherein said first reagent comprises a lipoprotein solubilization agent, a lipoprotein interactant, and a buffer;
adding to said first combined solution a second reagent to provide a second combined solution, wherein said second reagent comprises a buffer, an amphiphilic agent, a cholesterol esterase, a cholesterol oxidase, and a colorant, wherein said time of addition of said second reagent to said first combined solution is termed the initial time;
and
measuring absorbance of light by said second combined solution within a first time period, within a second time period, and after a third time, wherein said time periods and said third time may be determined with respect to said initial time, and
measuring the lipoprotein amounts of at least two lipoprotein components in the blood sample from said absorbance measurements,
whereby at least two lipoprotein components in a blood sample from a subject are measured.

3 more elements

A method for the combined measurement of high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL-C), total cholesterol (TC), and triglyceride level (TG) in no more than two samples of blood from a subject, said method comprising:

combining at least a portion of a first sample or first portion of a sample of blood from said subject with a first reagent comprising a lipoprotein solubilization agent, a lipoprotein interactant, and a buffer to provide a first combined solution;
adding to said first combined solution a second reagent comprising a buffer, an amphiphilic agent, a cholesterol esterase, a cholesterol oxidase, and a colorant to provide a second combined solution, wherein said time of addition of said second reagent to said first combined solution is termed the initial time;
and
measuring absorbance of light by said second combined solution within a first time period, within a second time period, and at a third time, wherein said time periods and said third time may be determined with respect to said initial time,
wherein HDL-C is measured by the difference in ΔA measured at said initial time and the ΔA measured at the end of said first time period, where ΔA is the difference between absorbance measured at two different wavelengths of light;
wherein LDL-C is measured by the difference in ΔA measured at the beginning of said second time period and the ΔA measured at the end of said second time period;
wherein TC is measured by ΔA measured at said third time;
whereby measurement of HDL-C, LDL-C, and TC is performed on said first sample of blood or said first portion of said sample of blood;
measuring TG in a second sample of blood or a second portion of a sample of blood from said subject;
and
calculating VLDL-C using the results of said TC, HDL-C, and TG measurements;
whereby combined measurement of HDL-C, LDL-C, VLDL-C, TC, and TG in two samples of blood or two portions of a sample of blood is obtained.

9 more elements

Dependencies are inferred from the claim text.

Rapid, low-sample-volume cholesterol and triglyceride assays

Life of this patent

Rapid, low-sample-volume cholesterol and triglyceride assays

Life of this patent

Abstract

Claims27