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Seven Direct Methods for Measuring HDL and LDL Cholesterol Compared with Ultracentrifugation Reference Measurement Procedures
Open accessAbstract
Background: Methods from 7 manufacturers and 1 distributor for directly measuring HDL cholesterol (C) and LDL-C were evaluated for imprecision, trueness, total error, and specificity in nonfrozen serum samples. Methods: We performed each direct method according to the manufacturer’s instructions, using a Roche/Hitachi 917 analyzer, and compared the results with those obtained with reference measurement procedures for HDL-C and LDL-C. Imprecision was estimated for 35 runs performed with frozen pooled serum specimens and triplicate measurements on each individual sample. Sera from 37 individuals without disease and 138 with disease (primarily dyslipidemic and cardiovascular) were measured by each method. Trueness and total error were evaluated from the difference between the direct methods and reference measurement procedures. Specificity was evaluated from the dispersion in differences observed. Results: Imprecision data based on 4 frozen serum pools showed total CVs <3.7% for HDL-C and <4.4% for LDL-C. Bias for the nondiseased group ranged from −5.4% to 4.8% for HDL-C and from −6.8% to 1.1% for LDL-C, and for the diseased group from −8.6% to 8.8% for HDL-C and from −11.8% to 4.1% for LDL-C. Total error for the nondiseased group ranged from −13.4% to 13.6% for HDL-C and from −13.3% to 13.5% for LDL-C, and for the diseased group from −19.8% to 36.3% for HDL-C and from −26.6% to 31.9% for LDL-C. Conclusions: Six of 8 HDL-C and 5 of 8 LDL-C direct methods met the National Cholesterol Education Program total error goals for nondiseased individuals. All the methods failed to meet these goals for diseased individuals, however, because of lack of specificity toward abnormal lipoproteins.
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“Most discordances in dyslipidemic samples are observed at lower concentration ranges of LDLC (< 1.8 mmol/L) and HDLC (< 1.0 mmol/L) [42].”
From Quantifying atherogenic lipoproteins for lipid-lowering strategies: Consensus-based recommendations from EAS and EFLM (2020) · cites this paper as [42]
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In the citing paper
“Direct measurements of dLDLC and dHDLC in normolipidemic samples usually meet the National Cholesterol Education Program (NCEP) total error goals of ≤12% and ≤13%, respectively, but total error ranged from -26% to +32% for dLDLC and -20% to +36% for dHDLC in a comprehensive study of different assays in dyslipidemic samples [42].”
From Quantifying atherogenic lipoproteins for lipid-lowering strategies: Consensus-based recommendations from EAS and EFLM (2020) · cites this paper as [42]
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“It is important to note that direct LDL-C measurements also have limitations, including systematic bias and inaccuracy in patients with dyslipidaemia, especially for high TG levels [88-90].”
From 2019 ESC/EAS guidelines for the management of dyslipidaemias: lipid modification to reduce cardiovascular risk (2019) · cites this paper as [89]
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Martin’s, Friedewald’s and Cordova’s formulas compared to LDL-C directly measured in Southern Brazil
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InfluentialCardiovascular disease lipids and lipoproteins biomarker standardization
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“Even in healthy individuals, calculated LDL-C may vary by 13.3–13.5% 7 .”
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Is data mining approach a best fit formula for estimation of low-density lipoprotein cholesterol?
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BackgroundCalculated values of serum LDL-cholesterol (LDL-C) – for better or worse?
Ramasamy, J., Job, V., Mani, T., & Jacob, M. (2021) · Nutrition Metabolism and Cardiovascular Diseases · doi:10.1016/j.numecd.2021.01.016 · Cited by 3
Tan, H. T., Yong, S., Liu, H., Liu, Q., Teo, T. L., & Sethi, S. K. (2023) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2022-1301 · Cited by 3
The necessity for improving lipid testing reagents: A real world study
Zheng, C., Zhou, W., Zhou, R., Zhang, H., Liang, S., Zhang, W., Li, G., Zhang, C., & Gan, W. (2023) · Clinica Chimica Acta · doi:10.1016/j.cca.2023.117529 · Cited by 3
Kilpatrick, E. S., Kallner, A., Atkin, S. L., & Sathyapalan, T. (2024) · Annals of Clinical Biochemistry International Journal of Laboratory Medicine · doi:10.1177/00045632241305936 · Cited by 3
Gupta, H., Schiros, C. G., Sharifov, O. F., Jain, A., & Denney, T. S. (2016) · BMC Cardiovascular Disorders · doi:10.1186/s12872-016-0352-x · Cited by 3
“These operating characteristics may not hold true for many commercial assays [23].”
BackgroundCalculated low density lipoprotein cholesterol: Its relevance in Indian perspective
Nigam, P. K. (2017) · Indian Heart Journal · doi:10.1016/j.ihj.2017.01.007 · Cited by 3
“Fourthly, for the equations incorporating HDLc the procedure for HDLc(direct/precipitation) should also be taken into consideration as bias have been reported even in the directly assayedHDLc values by different direct assays as compared to reference HDLc values.(4,15)”
ResultOhba, K., Oki, Y., Fujita, K., Kameko, F., Moriyama, T., Horiike, Y., Morita, H., Matsushita, A., Iino, K., Sasaki, S., et al. (2013) · Clinica Chimica Acta · doi:10.1016/j.cca.2013.09.003 · Cited by 3
Gerique, J. A. G., Ballesteros, B. M., Sardina, R. G., Salán, M. E., Romero, F. F., Puzo, J., Doreste, J. Á. A., Calmarza, P., Vázquez, M. D., & Romero, C. (2011) · Revista del Laboratorio Clínico · doi:10.1016/j.labcli.2011.09.005 · Cited by 3
Immunoprecipitation of apolipoprotein B-containing lipoproteins for isolation of HDL particles
Contois, J. H., Albert, A. L., & Nguyen, R.-A. (2014) · Clinica Chimica Acta · doi:10.1016/j.cca.2014.06.017 · Cited by 3
Seki, R., Inoue, K., Yamamoto, S., & Akimoto, K. (2017) · Biomedical Research and Clinical Practice · doi:10.15761/brcp.1000141 · Cited by 3
“Further, direct homogeneous methods have been shown to yield inconsistent data when IDL is elevated [28].”
BackgroundEngineering of a miniaturized, robotic clinical laboratory
Nourse, M. B., Engel, K., Anekal, S. G., Bailey, J. A., Bhatta, P., Bhave, D. P., Chandrasekaran, S., Chen, Y., Chow, S., Das, U., et al. (2017) · Bioengineering & Translational Medicine · doi:10.1002/btm2.10084 · Cited by 3
Pallazola, V. A., Quispe, R., Elshazly, M. B., Vakil, R. M., Sathiyakumar, V., Jones, S. R., & Martin, S. S. (2018) · Current Cardiovascular Risk Reports · doi:10.1007/s12170-018-0590-9 · Cited by 3
Agha, A. M., & Ballantyne, C. M. (2022) · Contemporary cardiology · doi:10.1007/978-3-030-98824-1_4 · Cited by 3
A failure of standardization or a failure of the process of standardization
Sniderman, A. D., & McQueen, M. (2018) · Journal of clinical lipidology · doi:10.1016/j.jacl.2018.05.009 · Cited by 3
Calculating LDL cholesterol in familial combined hyperlipidemia: Out with the old, in with the new?
Martin, S. S. (2018) · Atherosclerosis · doi:10.1016/j.atherosclerosis.2018.07.034 · Cited by 3
Lee, J. S., Lee, H., Cha, H., Seok, J., & Jeong, I. C. (2025) · PLoS ONE · doi:10.1371/journal.pone.0327169 · Cited by 2
“Although a previous study [39] reported that the Sekisui assay failed to meet the NCEP–ATP III total allowable error (TAE) criterion of ±12% in certain diseased populations, more recent evidence from a multicenter study by Miida et al. [40] demonstrated that the assay satisfied the NCEP–ATP III…”
Kathariya, G., Aggarwal, J., Nandal, R., & Reddy, S. (2025) · Heart India · doi:10.4103/heartindia.heartindia_77_25 · Cited by 2
Kuklenyik, Z., Ivanova, A. A., Drinkard, L. E., Schieltz, D. M., Jones, J. I., Bierbaum, K., Toth, C. A., Gardner, M. S., Parks, B. A., Andrews, M., et al. (2026) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-026-06390-9 · Cited by 2
Meikle, T. G., Wu, J., Wang, T., Dakic, A., Yi, C., Mellett, N. A., van Buuren-Milne, M., Beyene, H. B., Cinel, M. A., Duong, T., et al. (2026) · Analytical Chemistry · doi:10.1021/acs.analchem.5c04984 · Cited by 2
Diederiks, N. M., Romijn, F. P. H. T. M., van Neer, N. J. M., Pieterse, M., Dittrich, J., Ceglarek, U., Cobbaert, C. M., Ruhaak, L. R., the IFCC Working Group for Standardization of Apolipoproteins by Mass Spectrometry, Ruhaak, L. R., et al. (2026) · Clinical Chemistry · doi:10.1093/clinchem/hvag041 · Cited by 2
Non-HDL Estimation Methods: Advancing Cardiovascular Disease Prediction
Akbar, A. M., Jha, D., Ali, H., & Ahmad, T. (2023) · Indian Journal of Pharmacy Practice · doi:10.5530/ijopp.16.4.52 · Cited by 2
Which LDL Value Should Clinicians Look at?
Malve, H., & Asalkar, A. (2021) · Journal of Laboratory Physicians · doi:10.1055/s-0041-1729131 · Cited by 2
Lipid and Lipoprotein Abnormalities in Chronic Renal Insufficiency: Review
Rácz, O., Gaško, R., & Klimov, E. (2011) · InTech eBooks · doi:10.5772/22122 · Cited by 2
“Another unsolved problem is that the assay methods from different providers are not yet harmonized at all (Fuentes-Arderiu et al, 2009, Miller et al, 2010).”
BackgroundThe New ACC/AHA Cardiovascular Risk Guidelines: Impact and Controversies
Remaley, A. T., Shamburek, R. D., & Amar, M. J. (2014) · Clinical Chemistry · doi:10.1373/clinchem.2014.224964 · Cited by 2
Self-citationA Critical Review of LDL Cholesterol and HDL Cholesterol Measurement
Contois, J. H. (2012) · Cited by 2
Prevalence of Elevated Serum Homocysteine and Serum Lipoprotein ‘a’ in Women
Dhat, V. (2014) · JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH · doi:10.7860/jcdr/2014/9607.4995 · Cited by 2
Le point sur les différentes méthodes pour déterminer le cholestérol-LDL
Duvillard, L. (2011) · Médecine des Maladies Métaboliques · doi:10.1016/s1957-2557(11)70277-2 · Cited by 2
Solnica, B., Sygitowicz, G., Sitkiewicz, D., Cybulska, B., Jozwiak, J. J., Odrowąż‐Sypniewska, G., & Banach, M. (2020) · Diagnostyka Laboratoryjna · doi:10.5604/01.3001.0014.1296 · Cited by 2
Introduction of HDL Molecules, Past and Brief Future
Tabuchi, M., & Komoda, T. (2013) · Elsevier eBooks · doi:10.1016/b978-0-12-407867-3.00001-9 · Cited by 1
Michenaud, L., Marrié, N., Rimbert, A., Marmontel, O., Charrière, S., Gibert, C., Bouveyron, C., Mammi, J., Cariou, B., Moulin, P., et al. (2024) · Clinical Chemistry and Laboratory Medicine (CCLM) · doi:10.1515/cclm-2024-0587 · Cited by 2
Ercan, Ş., & Manav, A. (2025) · Biochemia Medica · doi:10.11613/bm.2025.020704 · Cited by 3
Kuti, M. A., Adeleye, J. O., Akinyemi, J. O., Ogundeji, O. A., Omoyele, O. O., Obe, O. A., Adewoyin, A. S., & Soriyan, O. O. (2025) · African Journal of Laboratory Medicine · doi:10.4102/ajlm.v14i1.2729 · Cited by 1
“4,5 There are, however, challenges with the use of the FF. Firstly, its inability to accurately estimate LDL cholesterol when the triglycerides are ≥ 4.52 mmol/L means that the laboratory needs to have an alternative means of determining and reporting LDL cholesterol in such scenarios.”
Influential
Metadata
- Authors
- Miller, W. G., Myers, G. L., Sakurabayashi, I., Bachmann, L. M., Caudill, S. P., Dziekonski, A., Edwards, S., Kimberly, M. M., Korzun, W. J., Leary, E. T., Nakajima, K., Nakamura, M., Nilsson, G., Shamburek, R. D., Vetrovec, G. W., Warnick, G. R., & Remaley, A. T.
- Affiliations
- Virginia Commonwealth University (US)
- Centers for Disease Control and Prevention (US)
- Jichi Medical University (JP)
- Virginia Commonwealth University, Richmond, VA
- Pacific Biomarkers (United States) (US)
- Otsuka (Japan) (JP)
- Osaka Medical Center for Health Science and Promotion, Osaka, Japan
- Nilsson Measurement Quality, Uppsala, Sweden
- National Institutes of Health (US)
- Health Diagnostics Laboratory, Richmond, VA
- Corresponding author
- Miller, W Greg
- ORCID
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2010-06-01
- Volume
- 56
- Issue
- 6
- Pages
- 977-986
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- PMC
- PMC4687457
- OpenAlex
- W2130132335
- Cited by
- 386
- References
- 23
- Citations by year
- 2026: 22 · 2025: 31 · 2024: 25 · 2023: 31 · 2022: 30 · 2021: 30 · 2020: 31 · 2019: 26 · 2018: 22 · 2017: 21 · 2016: 15 · 2015: 14 · 2014: 27 · 2013: 16 · 2012: 17
- FWCI
- 13.22
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Clinical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Merck
- Pfizer
- Abbott
- NIH
- Topics
- Health Sciences › Health Professions › Medical Laboratory Technology › Clinical Laboratory Practices and Quality Control
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Lipoproteins and Cardiovascular Health
- Physical Sciences › Physics and Astronomy › Instrumentation › Scientific Measurement and Uncertainty Evaluation
- Keywords
- HDL cholesterol, LDL cholesterol, direct measurement method, method comparison, total error, imprecision, trueness, specificity, dyslipidemia, cardiovascular disease
- MeSH terms
- Blood Chemical Analysis, Humans, Cholesterol, HDL, Cholesterol, LDL, Reference Standards, Sensitivity and Specificity, Ultracentrifugation
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