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Clinical Chemistry 44: 1964-1973, 1998;
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(Clinical Chemistry. 1998;44:1964-1973.)
© 1998 American Association for Clinical Chemistry, Inc.

General Clinical Chemistry

Luminometric single step urea assay using ATP-hydrolyzing urease

Birgitta Näslunda, Lars Ståhle, Arne Lundin1, Björn Anderstam, Peter Arner, and Jonas Bergström

1 BioThema AB, Strandvägen 36, S-130 54 Dalarö, Sweden.
a Address correspondence to this author at: Department of Pharma Marketing, Roche AB, Liljeholmsstranden 5, Box 47327, S-100 74 Stockholm, Sweden. Fax 46-87440681; e-mail birgitta.naslund{at}roche.com.

An automatic enzyme kinetic luminometric method for determination of small quantities of urea in biological fluids and in microdialysates is presented. The method is based on the ATP-hydrolyzing urease reaction [urea amidohydrolase (ATP-hydrolyzing); EC 3.5.1.45], monitored by a luciferin-luciferase ATP reaction. The assay range is 100 pmol to 50 nmol with a detection limit of 5 µmol/L in the sample, compared with detection limits of 0.1 mmol/L in earlier spectrophotometric methods. To reduce the non-urea-dependent ATPase activity (vblank) and to increase the urea-dependent activity, 1,2-propanediol was included. Assay conditions were optimized by multivariate analysis. Recoveries of urea added to blood dialysate and plasma were 96–103%. No analytical interference of common metabolites, drugs, or other additives was observed. The total CVs (6 days and six concentrations, 1.2–21.8 mmol/L) were 3.6–8.5%. The results obtained with the present assay were highly correlated for dialysate (r = 0.979) and for plasma (r = 0.978) with those obtained by a spectrophotometric kit method with slopes of 1.02–1.03 and intercepts of 0.08–0.23 mmol/L.




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


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 Am. J. Pathol.Home page
M. A. Olszewski, M. C. Noverr, G.-H. Chen, G. B. Toews, G. M. Cox, J. R. Perfect, and G. B. Huffnagle
Urease Expression by Cryptococcus neoformans Promotes Microvascular Sequestration, Thereby Enhancing Central Nervous System Invasion
Am. J. Pathol., May 1, 2004; 164(5): 1761 - 1771.
[Abstract] [Full Text] [PDF]


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