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Published on: October 12, 2017
Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects
Takashi Miida1, Kunihiro Nishimura2, Tomonori Okamura3
1Department of Clinical Laboratory Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
Insights
Homogeneous assays for high-density lipoprotein-cholesterol (HDL-C) generally agree with reference methods in patients, though accuracy varies in those with very low HDL-C or abnormal composition.
Area of Science:
- Clinical Chemistry
- Cardiovascular Risk Assessment
- Biomarker Validation
Background:
- High-density lipoprotein-cholesterol (HDL-C) is a key negative risk factor for cardiovascular events.
- Accurate measurement of HDL-C is crucial for risk stratification.
- Validation of available homogeneous HDL-C assays is needed, especially in diverse patient populations.
Purpose of the Study:
- To evaluate the agreement between homogeneous HDL-C assays and reference measurement procedures (RMPs).
- To assess assay accuracy in both healthy individuals and subjects with various diseases.
Main Methods:
- Compared HDL-C concentrations from 12 homogeneous assays against RMPs (ultracentrifugation/heparin-manganese precipitation).
- Analyzed fresh clinical samples from 48 healthy subjects and 119 subjects with disease.
- Assessed assay performance using bias, total error, and correlation analyses.
Main Results:
- Most homogeneous assays demonstrated excellent precision (CVs <2.23%).
- Nine reagents met National Cholesterol Education Program total error requirements in healthy subjects; six met them in subjects with disease.
- Agreement with RMPs was generally good, with most biases within ±1.0%, but some reagents showed significant errors in specific patient groups.
Conclusions:
- Homogeneous HDL-C assays largely agree with reference methods in subjects with common diseases.
- Accuracy may be compromised in individuals with extremely low HDL-C levels or abnormal HDL particle composition.
- Three specific reagents showed notable discrepancies requiring further investigation.
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.
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