Rapid, low-sample-volume cholesterol and triglyceride assays

摘要

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.

主权项

1. 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 of said second combined solution at two different wavelengths 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.