Adenosine Deaminase Assay
ADA is an enzyme catalyzing the deamination reaction from adenosine to inosine. The enzyme is widely distributed in human tissues, especially high in T lymphocytes. Elevated serum ADA activity has been observed in patients with acute hepatitis, alcoholic hepatic fibrosis, chronic active hepatitis, liver cirrhosis, viral hepatitis, and hepatoma. Increased ADA activity was also observed in patients with tuberculous effusions. Determination of ADA activity in patient serum may add unique values to the diagnosis of liver diseases in combination with ALT or γ-GT (GGT) tests. ADA assay may also be useful in the diagnostics of tuberculous pleuritis.
The ADA assay is based on the enzymatic deamination of adenosine to inosine which is converted to hypoxanthine by purine nu-cleoside phosphorylase (PNP). Hypoxanthine is then converted to uric acid and hydrogen peroxide (H2O2) by xanthine oxidase (XOD). H2O2 is further reacted with N-Ethyl-N-(2hydroxy-3-sulfopropyl)-3-methylaniline (EHSPT) and 4aminoantipyrine (4-AA) in the presence of peroxidase (POD) to generate quinone dye which is monitored in a kinetic manner. The entire enzymatic reaction scheme is shown below.
Adenosine + H2O → Inosine + NH3
Inosine + Pi → Hypoxanthine + Ribose-1-phosphate
Hypoxanthine +2H2O + 2O2 → Uric acid + 2H2O2
4H2O2 + 4-AA + EHSPT → 4H2O +Quinone dye (λ max 556nm)
One unit of ADA is defined as the amount of ADA that generates one µmole of inosine from adenosine per min at 37°C.
- Kobayashi F, Ikeda T, Marumo F, Sato C: Adenosine deaminase isoenzymes in liver disease. Am. J. Gastroenterol. 88: 266-271 (1993)
- Kallkan A., Bult V., Erel O., Avci S., and Bingol N. K.: Adenosine deaminase and guanosine deaminase activities in sera of patients with viral hepatitis. Mem Inst. Oswaldo Cruz 94(3) 383386 (1999)
- Burgess LJ, Maritz FJ, Le Roux I, et al. Use of adenosine deaminase as a diagnositic tool for tuberculous pleurisy. Thorax 50: 672-674 (1995)