Maria Cristina De Martino

Validação laboratorial de um método automatizado de dosagem da atividade de adenosina desaminase em líquido pleural e em líquido cefalorraquidiano

OBJECTIVE The incidence of tuberculosis worldwide has emphasized the need for better assays designed to diagnose the disease, principally the extrapulmonary form. The objective of the present study was to validate the performance of an automated method for the determination of adenosine deaminase (ADA) activity in pleural fluid (PF) and cerebrospinal fluid (CSF), comparing it with a conventional method (the modified Giusti method). METHODS In total, 134 samples were selected from among those tested in our laboratory: 94 PF samples and 40 CSF samples. The ADA activity was determined using the two methods. Inter- and intra-assay precision was determined, linear regression analysis was performed, simple concordance tests were conducted, and the means of the differences were calculated. RESULTS The correlation coefficients for PF and CSF samples were, respectively, 0.96 and 0.95. Inter-assay precision was determined using 21 replicates at 3 different activity levels: low, medium and high. The percentage coefficient of variation (%CV) was, respectively, 5.9, 8.1 and 5.8 for PF samples, compared with 21.9, 18.6 and 13.8 for CSF samples. Intra-assay precision in %CV was 1.3 and 11.7, respectively, for PF and CSF samples. The concordance between the methods in PF and CRF samples was, respectively, 96.8% and 100%, considering the reference values for the diagnosis of TB to be 40 U/L (conventional) and 30 U/L (automated) in PF samples, versus 9 U/L (for both methods) in CSF samples. CONCLUSIONS The results validate the use of the automated method of determining ADA activity in PF and CSF samples as an alternative to the conventional method.

Implications for the use of acid preservatives in 24-hour urine for measurements of high demand biochemical analytes in clinical laboratories.

BACKGROUND Evaluate the level of interference of biochemists dosages in the 24-hour urine using or not the 6 mol/l HCl acid in different concentrations and conditions and its implications in the most demanded analytes in clinical laboratory. METHODS Twenty-two volunteers collected three 24-hour urine in 3 conditions: with 5 ml/l and 20 ml/l of 6 mol/l HCl in the container, and without acid preservative. The samples collected without preservative were separated in aliquots and added 5 ml/l of 6 mol/l HCl after 24 h. Analytes, uric acid creatinine, urea, chlorides, glucose, magnesium, sodium, potassium, microalbumin, proteins, amylase, aldosterone, calcium, cortisol, phosphorus, citric acid, oxalate, and metanephrines, were determined. RESULTS Uric acid, glucose, microalbumin, protein, amylase and aldosterone showed that %CV ranging from 16 to 57% in the presence of acid preservative. Analytes that need acid preservative cortisol, citric acid and oxalate showed %CV ranging from 6 to 27% with r=0.66, r=0.77, r=0.70 respectively provided 5 ml/l after delivery and r=0.31, r=0.70 and r=0.48 without preservative acid when compared with the gold standard (with 20 ml/l of 6 mol/l HCl). CONCLUSIONS Glucose, microalbumin, protein, amylase and aldosterone urinary did not show good performance in the presence of acid preservative. Analytes that need acid preservative showed variation acceptable in condition 5 ml/l of 6 mol/l HCl added after 24 h.