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Clinical Chemistry 49: 1105-1113, 2003; 10.1373/49.7.1105
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(Clinical Chemistry. 2003;49:1105-1113.)
© 2003 American Association for Clinical Chemistry, Inc.

Time-resolved Fluorescence Resonance Energy Transfer Assay for Point-of-Care Testing of Urinary Albumin

Qiu-Ping Qin1,a, Olli Peltola2 and Kim Pettersson1

1 Department of Biotechnology, University of Turku, Tykistökatu 6, 20520 Turku, Finland.

2 Central Laboratory of Turku University Hospital, Kiinamyllynkatu 4-8, 20521 Turku, Finland.

aAuthor for correspondence. Fax 358-2-3338050; e-mail qiqin{at}utu.fi.

Background: Microalbuminuria is an established early marker of diabetic nephropathy and an important cardiovascular risk factor in diabetes and hypertension. We aimed to develop a rapid point-of-care assay for the measurement of urine albumin.

Methods: The competitive homogeneous assay used an albumin-specific monoclonal antibody labeled with a stable fluorescent europium chelate as donor and an albumin labeled with cyanine 5 (Cy5) as acceptor. The assay was performed at room temperature in single microtitration wells that contained all the required dry-form reagents. The close proximity between the two labels in the immune complex allowed fluorescence resonance energy to be transferred from the pulse-excited europium chelate to the acceptor Cy5. The emission of long-lived energy transfer signal from the sensitized Cy5 was measured at 665 nm with time-resolved fluorometry that eliminated short-lived background.

Results: The assay procedure required 12 min for a 10-µL urine sample. The working range was from 10 to ~320 mg/L, and the lower limit of detection was 5.5 mg/L. The within- and between-run CVs were 6.9–10% and 7.5–13%, respectively. Recovery was 103–122%. The assay correlated well (r2 = 0.98; n = 37) with a laboratory-based immunoassay, although mean (SD) results were 7 (29)% lower.

Conclusions: The speed and ease of performance of this assay recommend it for near-patient use. The assay is the first to combine a fluorescence resonance energy transfer-type rapid competitive assay with an all-in-one dry reagent.




The following articles in journals at HighWire Press have cited this article:


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[Abstract] [Full Text] [PDF]


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