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Agreement among Four Homocysteine Assays and Results in Patients with Coronary Atherosclerosis and Controls

¹ Departments of Laboratory Medicine and Pathology, Children’s Hospital, Harvard Medical School, Boston, MA 02115.
Departments of
² Clinical Laboratories and
³ Cardiology, University of Damascus Hospitals, Damascus, Syria.

⁴ Department of Biochemistry and Microbiology, School of Pharmacy, University of Damascus, Damascus, Syria.
^a Address correspondence to this author at: Children’s Hospital, Department of Laboratory Medicine, 300 Longwood Ave., Boston, MA 02115. E-mail rifai{at}a1.tch.harvard.edu

Background: Hyperhomocysteinemia has been associated with coronary atherosclerosis in many, but not all, prospective and retrospective studies. Some on these inconsistencies may be attributed to methodological variabilities.

Methods: In the present study, three newly commercially available assays and one in-house HPLC assay for total homocysteine (tHcy) were utilized in 99 subjects with angiographically documented atherosclerosis and in 91 community controls matched by age, gender, and smoking history. The in-house assay, a modified Fortin and Genest HPLC method, was compared with the Bio-Rad HPLC assay, the Abbott IMx^® fluorescence polarization immunoassay, and a Bio-Rad enzyme-linked immunoassay (EIA) microtiter method.

Results: Correlation coefficient values between the in-house HPLC assay and the Bio-Rad HPLC, the Abbott IMx, and the Bio-Rad EIA assays were 0.95, 0.96 and 0.90, respectively. Although tHcy concentrations were higher in cases compared with controls by all four methods, the difference reached statistical significance only with the in-house HPLC procedure (median, 13.5 ± 6.7 µmol/L in cases vs 10.9 ± 4.8 µmol/L in controls; P <0.01, adjusting for covariates), where it was an independent predictor of case or control status, along with hypertension, total cholesterol, and triglycerides. The tHcy distributions in cases and controls demonstrated significant overlap. The number of atherosclerotic major coronary vessels was associated with significantly higher tHcy (P <0.01 for trend) in all four methods.

Conclusions: The three commercial assays for tHcy differed in analytical and clinical performance. Analytically, the Abbott IMx method showed the best comparability with the in-house assay, but clinically, the three commercial methods were similar and did not distinguish cases from controls.

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