Validation of a novel homogeneous assay for of HDL3-C measurement

• Published in: Clinica chimica acta Vol. 425; pp. 37 - 41
• Main Authors: Ashmaig, M.E., Gupta, S., McConnell, J.P., Warnick, G.R.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 21.10.2013
• Subjects: Bilirubin - blood; Calibration; Chemical Precipitation; Cholesterol, HDL - blood; Cholesterol, HDL - classification; Colorimetry - methods; Colorimetry - standards; CVD; Dextran Sulfate - chemistry; HDL-C; HDL-subclasses; HDL2-C; HDL3-C; Hemoglobins - metabolism; Humans; Reproducibility of Results; Sensitivity and Specificity; Triglycerides - blood; HDL-C; CVD; HDL-subclasses; HDL2-C; HDL3-C
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/j.cca.2013.07.007

Low serum concentration of high density lipoprotein2 cholesterol (HDL2-C) is associated with increased risk of cardiovascular events. HDL2-C is calculated indirectly by subtracting high density lipoprotein3 cholesterol (HDL3-C) from total high density lipoprotein cholesterol (HDL-C). However, the special equipment and long assay times required for HDL3-C measurement have hindered the use of HDL2-C clinically. Here, we report the validation of a simple and rapid homogeneous assay for HDL3-C that is adaptable to clinical chemistry analyzers. Method comparison based on 2740 serum specimens spanning the physiological range of HDL3-C was analyzed in singlicate to evaluate and validate a new homogeneous assay from Denka Seiken against the conventional dextran sulfate precipitation method. This study was performed over five days. Serum pools were prepared for the analysis of precision over 5days (5 measurements per day), linearity, and interference (hemoglobin, bilirubin, and triglycerides) evaluation. The homogeneous method had good within-run precision at concentrations of 24, 36, and 46mg/dl, yielding standard deviations (SD) of 0.2 (0.9%) 0.4 (1.2%), and 0.5 (1.1%), respectively. Between-day precision, performed over 5days using the same serum pools, yielded SD of 0.3 (1.4%), 1.0 (2.8%), and 0.9 (2.0%), respectively. The assay was linear from 1 to 100mg/dl and correlated very well with the dextran sulfate precipitation method. There was no interference from hemoglobin up to 500mg/dl, bilirubin up to 25mg/dl, or triglycerides up to 1500mg/dl. This homogeneous HDL3-C assay quantitatively measures HDL3-C in serum samples and has excellent precision, and can be implemented on an automated chemistry analyzer, thereby facilitating rapid measurement (~10min) of a large number of samples in a standard clinical laboratory without the need for additional expensive equipment, laboratory space, or specially-trained staff. •First homogeneous HDL3-C assay that quantitatively measures HDL3-C in serum•Calculation of HDL2-C by subtracting HDL3-C from total HDL-C•Adaptable to automated chemistry analyzer for rapid measurement of HDL3-C in serum•Most efficient and cost effective method used for HDL subclasses measurements