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https://doi.org/10.11613/BM.2020.030703

Analysis of serum homocysteine in the laboratory practice - comparison of the direct chemiluminescence immunoassay and high performance liquid chromatography coupled with fluorescent detection

Łukasz Paprotny ; Research and Development Center, Alab Laboratories, Lublin, Poland
Dorota Wianowska ; Department of Chromatographic Methods, Faculty of Chemistry, Maria Curie-Skłodowska University, Lublin, Poland
Magdalena Izdebska ; Department of Laboratory Diagnostics, Medical University of Lublin, Lublin, Poland
Agnieszka Celejewska ; Research and Development Center, Alab Laboratories, Lublin, Poland
Dorota Szewczak ; Research and Development Center, Alab Laboratories, Lublin, Poland
Janusz Solski ; Department of Laboratory Diagnostics, Medical University of Lublin, Lublin, Poland

Sažetak

Introduction: Effective diagnosis of cardiovascular diseases requires the right tools to be used enabling selective and sensitive analysis of their biomarkers.
One of them is homocysteine (Hcy), nowadays determined by immunoassays and chromatographic methods. This study aims to compare
the results obtained by direct chemiluminescence immunoassay (CLIA) and high performance liquid chromatography with fluorescent detection
(HPLC-FD) using commercial kits.
Materials and methods: Homocysteine concentration was determined in serum samples obtained from 101 individuals, using Atellica IM HCY
(Siemens Healthineers, Erlangen, Germany) and HCY in plasma/serum – HPLC-FD (Chromsystems Instruments & Chemicals GmbH, Gräfelfing, Germany)
tests validated for routine analysis. The latter was applied as a reference method. The comparability and agreement between the tested methods
were evaluated using the Passing-Bablok (PB) regression analysis and the Bland-Altman (BA) method of the differences analysis.
Results: Studies showed that CLIA gives higher Hcy concentrations (15.7 ± 4.14 μmol/L). Passing-Bablok regression analysis of the results obtained
with CLIA (y) compared with HPLC-FD (x) yielded an intercept of 0.22 (95%CI: - 2.16 to 2.46) and slope of 1.58 (95%CI: 1.33 to 1.87). Bland-Altman
analysis demonstrated a systematic positive bias for CLIA of 5.85 ± 2.77 μmol/L.
Conclusions: Methods disagreement precludes their interchangeability. Lower Hcy values by HPLC-FD result from its greater selectivity. High performance
liquid chromatography with fluorescent detection should be considered as preferential method for analysing Hcy in blood serum as well as
the recommended reference method for routine clinical analysis. This fact, however, imposes the need to establish new reference ranges.

Ključne riječi

homocysteine; immunoassay; luminescent measurements; liquid chromatography

Hrčak ID:

244727

URI

https://hrcak.srce.hr/244727

Datum izdavanja:

15.10.2020.

Posjeta: 751 *