Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects.
BACKGROUND
High-density lipoprotein-cholesterol (HDL-C) is a negative risk factor for cardiovascular events. Although several homogeneous HDL-C assays are available, their accuracy has not been validated, particularly in subjects with disease. We aimed to clarify whether HDL-C concentrations measured by homogeneous assays [HDL-C (H)] agree with those determined by the reference measurement procedures [HDL-C (RMP)] using ultracentrifugation and precipitation with heparin-manganese reagent in fresh clinical samples.


METHODS
HDL-C concentrations in samples from 48 healthy subjects and 119 subjects with disease were determined using 12 homogeneous assays and RMPs.


RESULTS
All reagents showed excellent intra- and inter-assay CVs (<2.23%) for two pooled sera. Furthermore, the mean bias was within ± 1.0% in nine reagents using samples from healthy subjects and in eight reagents using samples from subjects with disease. In a single HDL-C (H) determination, the total error requirement of the National Cholesterol Education Program (95% of results < 13%) was fulfilled in nine reagents using samples from healthy subjects and six reagents in those from subjects with disease. Error component analysis revealed that only one reagent exceeded ± 10% total error in samples from healthy subjects, whereas four reagents exceeded this error in samples from subjects with disease. Correlations between HDL-C (H) and HDL-C (RMP) revealed that the slopes were within 1.00 ± 0.06 in six reagents in healthy subjects, and eight reagents in subjects with disease.


CONCLUSIONS
Except for three reagents, HDL-C (H) agrees well with HDL-C (RMP) in subjects with common disease, but not in those with extremely low HDL-C or abnormal HDL composition.
['Miida|Takashi|T|', 'Nishimura|Kunihiro|K|', 'Okamura|Tomonori|T|', 'Hirayama|Satoshi|S|', 'Ohmura|Hirotoshi|H|', 'Yoshida|Hiroshi|H|', 'Miyashita|Yoh|Y|', 'Ai|Masumi|M|', 'Tanaka|Akira|A|', 'Sumino|Hiroyuki|H|', 'Murakami|Masami|M|', 'Inoue|Ikuo|I|', 'Kayamori|Yuzo|Y|', 'Nakamura|Masakazu|M|', 'Nobori|Tsutomu|T|', 'Miyazawa|Yukihisa|Y|', 'Teramoto|Tamio|T|', 'Yokoyama|Shinji|S|']
D015982:Bias D001774:Blood Chemical Analysis D008076:Cholesterol, HDL D004194:Disease D006801:Humans D007202:Indicators and Reagents D015203:Reproducibility of Results D014461:Ultracentrifugation
Designated comparison method Direct HDL-C assay Standardization