New immunoseparation-based homogeneous assay for HDL-cholesterol compared with three homogeneous and two heterogeneous methods for HDL-cholesterol.
We evaluated four new commercial methods for HDL-cholesterol determination. The three completely homogeneous assays were an immunoseparation-based (IS) method from Wako, a polyethylene glycol-modified enzyme (PEG) method from Boehringer Mannheim, and a synthetic polymer-based (SP) method from Genzyme. The fourth method was a new heterogeneous method in which lipoproteins are removed using dextran sulfate-coated magnetic beads and Mg2+ (MB, Reference Diagnostics). We compared these methods with the conventional phosphotungstic acid/MgCl2 precipitation (PTA) procedure. The homogeneous assays had good intraassay imprecision with total CVs <2.3%, whereas the CVs of the MB assay were <5.9%. Adding HDL to serum to achieve HDL-cholesterol (HDL-C) concentrations up to 1000 mg/L revealed nearly complete recoveries in the IS, PEG, and MB assays, whereas the SP assay showed a lower recovery (approximately 70%). The SP HDL-C apparently increased at increasing LDL-cholesterol and VLDL-triglyceride concentrations, whereas the IS, PEG, and MB methods were not influenced by LDL-cholesterol up to 6000 mg/L (MB, 5000 mg/L) and VLDL-triglycerides up to 9000 mg/L. Free fatty acids above approximately 2 mmol/L produced falsely high HDL-C in the IS and SP assays, the error amounting to as much as 50% in some samples. An intermethod comparison in 291 fresh serum samples yielded correlation coefficients of at least r = 0.95 for all assays, when compared with the PTA procedure. The slopes and intercepts of the regression lines were 1.05 and 57 (IS), 1.12 and 9.9 (PEG), 1.00 and 39 (SP), and 1.0 and 38 mg/L (MB), respectively. The new assays are precise and simplify the determination of HDL-C, but in part they lack specificity or are susceptible to interferences, resulting in discrepancies when compared with the established PTA procedure.
University Hospital of Freiburg, Germany. manauck@mz1200.ukl.uni-freiburg.de
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Wieland
D000906:Antibodies / Q000276:immunology; D001055:Apolipoproteins B / Q000276:immunology; D011232:Chemical Precipitation; D008076:Cholesterol, HDL / Q000097:blood*; D016264:Dextran Sulfate; D005230:Fatty Acids, Nonesterified / Q000097:blood; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007202:Indicators and Reagents; D015636:Magnesium Chloride; D008280:Magnetics; D010772:Phosphotungstic Acid; D011092:Polyethylene Glycols; D012680:Sensitivity and Specificity
D000906:Antibodies; D001055:Apolipoproteins B; D008076:Cholesterol, HDL; D005230:Fatty Acids, Nonesterified; D007202:Indicators and Reagents; D015636:Magnesium Chloride; D010772:Phosphotungstic Acid; D011092:Polyethylene Glycols; D016264:Dextran Sulfate