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Journal of Clinical Microbiology, April 2000, p. 1439-1443, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A Rapid, Automated Enzymatic Fluorometric Assay for Determination of D-Arabinitol in Serum

Siew Fah Yeo,1,2 Yeyan Zhang,1,2 David Schafer,3 Sheldon Campbell,3,4 and Brian Wong1,2,*

Departments of Internal Medicine1 and Laboratory Medicine,4 Yale University School of Medicine, New Haven, and Infectious Diseases2 and Laboratory Medicine and Pathology,3 Veterans Affairs Medical Center, West Haven, Connecticut

Received 20 September 1999/Returned for modification 23 December 1999/Accepted 2 February 2000

A rapid enzymatic fluorometric assay for measuring D-arabinitol in serum was developed using recombinant D-arabinitol dehydrogenase from Candida albicans (rArDH). rArDH was produced in Escherichia coli and purified by dye-ligand affinity chromatography. rArDH was highly specific for D-arabinitol, cross-reacting only with xylitol (4.9%) among all polyols tested. A Cobas Fara II centrifugal autoanalyzer (Roche) was used to measure NADH fluorometrically when rArDH and NAD were added to serum extracts, and D-arabinitol concentrations were calculated from standard curves derived from pooled human serum containing known amounts of D-arabinitol. The method was precise (mean intra-assay coefficients of variation [CVs], 0.8%, and mean interassay CVs, 1.6%) and rapid (3.5 min per assay) and showed excellent recovery of added D-arabinitol in serum (mean recovery rate, 101%). The mean and median D-arabinitol/creatinine ratios were 2.74 and 2.23 µM/mg/dl, respectively, for the 11 patients with candidemia compared to 1.14 and 1.23 µM/mg/dl, respectively, for 10 healthy controls (P < 0.01). These results confirm earlier studies showing that serum D-arabinitol measurement may help to promptly diagnose invasive candidiasis. The technique shows a significant improvement in terms of accuracy, cost, simplicity, specificity, and speed compared with gas chromatography, mass spectrometry, and earlier enzymatic assays.

* Corresponding author. Mailing address: VA Medical Center, Infectious Diseases, 950 Campbell Ave., 111-I, West Haven, CT 06516. Phone: (203) 932-5711, ext. 5168. Fax: (203) 937-4851. E-mail: brian.wong{at}yale.edu.

Journal of Clinical Microbiology, April 2000, p. 1439-1443, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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