Development of a rapid, microplate-based kinetic assay for measuring adenosine deaminase activity in body fluids

Clinica Chimica Acta; International Journal of Clinical Chemistry
Jun Lu, David G Grenache

Abstract

Adenosine deaminase (ADA) catalyzes the deamination of adenosine to inosine. The activity of ADA in body fluids has clinical utility in the assessment of suspected tuberculosis. The conversion of adenosine to inosine was monitored in 96-well microplates as a continuous decrease at 265 nm for 15 min at ambient temperature. Analytical precision, sensitivity, linearity, accuracy, and enzyme stability were validated. Reference intervals were established from >120 tuberculosis-negative pleural, peritoneal, and cerebrospinal fluid samples. The molar extinction coefficients of adenosine and inosine at 265 nm were 12,715 and 4,918 l/mol.cm and their difference was used to calculate ADA activity. Maximum within-day imprecision was <11% and maximum total precision was <19%. Analytical sensitivity was 0.5 U/l and the assay was linear to 40 U/l. ADA recovery was 96-110% over an activity range of 11.7-25.3 U/l. ADA was stable for 1, 7 and 30 days at ~25 °C, 4-8 °C, and -20 °C storage, respectively. Upper reference limits were 9.4, 7.3, and 1.5 U/l for pleural, peritoneal, and cerebrospinal fluid, respectively. The microplate-based kinetic ADA assay has favorable performance characteristics. This method eliminates the need for assay calibrat...Continue Reading

References

May 1, 1969·Annals of Human Genetics·D A HopkinsonH Harris
Nov 25, 2000·Clinical and Experimental Immunology·A Fischer
Oct 4, 2005·Alimentary Pharmacology & Therapeutics·F M Sanai, K I Bzeizi
Sep 21, 2010·Clinical Biochemistry·Hervé DelacourPascal Burnat

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Citations

Feb 8, 2013·Biomarkers in Medicine·Cyro Teixeira da SilvaElizabeth Giestal de Araújo
Oct 19, 2017·Applied Biochemistry and Biotechnology·Agnieszka Łupicka-SłowikMarcin Sieńczyk

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