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Indagine conoscitiva sulla diffusione delle analisi di biologia molecolare nei laboratori clinici della regione Puglia

Marina Micelli*, Mario Correale, Antonio Iacobacci, Sergio Giannattasio
*Laboratorio di Coagulazione-Azienda Ospedaliera Policlinico, 70124 Bari

Biochimica Clinica: 2001; 25(1): 5-9 [Article in italian]

ABSTRACT. In view of the increasing interest on the application of molecular biology techniques in the clinical laboratory, the need is felt for an appropiate knowledge of the existing laboratories performing molecular biology tests, in order to optimize a diagnotic service network in a region. Therefore, a survey has been organized in the Puglia region, by sending a questionnaire to 105 public clinical laboratories, asking them to give information about the molecular biology tests performed, the methododolgies, and the human and instrumental resources involved. Valid results were obtained from 75 participants: out of them, 35 declared to use molecolar biology techniques for the execution of at least one type of diagnostic test. The use of molecular biology techniques showed wider diffusion in the area of clinical microbiology, partially due to the availability of automatic instruments and diagnostic kits. The execution of 8530 HCV-RNA tests/year, of 3610 HIV-RNA tests year, and 3170 HBV-RNA tets/year, toghether with a number of additional microbiologic tests based on molecular biology, was recorded. In the field of hereditary and oncologic pathology the diffusion of molecular biology was much lower, and the number of tests per year was seldom higher than a few hundreds. The results from the survey are considered as a valid basis for future rationalization of the molecular biology and medical genetics services regional network.

Determinazione della lipoamide deidrogenasi nel siero: proposta di un nuovo approccio analitico-strumentale e confronto con altri indicatori di lesione mitocondriale

Cristina Serena*, Roberto Bonora, Mauro Panteghini
*Laboratorio Analisi Chimico Cliniche 1, Azienda Ospedaliera 'Spedali Civili', Brescia

Biochimica Clinica: 2001; 25(1): 10-16 [Article in italian]

ABSTRACT. The lipoamide dehydrogenase (LAD) is a mitochondrial flavoprotein which catalyzes the reduction of lipoamide by NADH to yield dihydrolipoamide and NAD+. The enzyme is a component of the multienzyme pyruvate dehydrogenase complex and 2-oxoglutarate dehydrogenase complex. LAD can be measured in human serum and used potentially as marker of tissue damage. In order to investigate the clinical utility of LAD determination, we adapted a modification of the manual method of Pelley et al. (Clin Chem 1976;22:275) for use on the Roche Cobas Bio analyzer. The assay measures the lipoamide-dependent reduction of NAD+ at 340 nm. The automated assay requires 15 mL of specimen and 140 mL of reagents containing NAD+ and reduced lipoamide in tricine buffer, 50 mmol/L. Following 10 min incubation, seven absorbance readings were taken at 10 s intervals (37°C). Under the conditions described, the products of reaction increased linearly with serum LAD activities up to 300 U/L. The average percentage recovery of various quantities of commercial LAD (Sigma) was 98.7% (CV 8.0%). Analysis of total imprecision, using NCCLS-EP5 protocol on three human sera with 15.6 U/L, 34.6 U/L, and 87.3 U/L LAD activities, gave CV of 15.6%, 7.8%, and 3.7%, respectively. Triglycerides, hemoglobin, and bilirubin do not interfere up to values of 500 mg/dL, 0.25 g/dL, and 10 mg/dL, respectively. Results of LAD obtained with our method (y) well correlated with those of manual method of Pelley (x) (r=0.9947) yielding the following regression equation: y = 1.02x + 0.92, n=23. The upper reference limit, determined in a group of 33 ostensibly healthy subjects, was 19 U/L (97.5th percentile). From presented data, we conclude that our automated method may now permit studies on the clinical utility of serum LAD.

Marcatori biochimici del metabolismo osseo in pazienti adulti con morbo celiaco

Giulio Mengozzi*, Cristina Baldi, Teresa Zaccaria, Silvia Bruna Grosso, Sandra Biasiol, Marco Di Stefano, Gian Carlo Isaia, Carla Sategna-Guidetti, Giuseppe Aimo, Roberto Pagni, Silvia Martini
*U.O.A. Laboratorio di Chimica Clinica "Baldi e Riberi", Azienda Ospedaliera San Giovanni Battista, Torino

Biochimica Clinica: 2001; 25(1): 17-22 [Article in italian]

ABSTRACT. The diagnostic accuracy and predictivity of biochemical markers of bone turnover as compared to bone mineral density (BMD) and the role of non-invasive tests in the characterization of bone metabolism derangement, with particular regard to osteomalacia, were evaluated in 86 consecutive newly diagnosed, biopsy confirmed, adult coelic patients. According to WHO criteria, 34% of patients had a normal BMD, 40% had osteopenia and 26% osteoporosis. Osteopenics had higher bone specific alkaline phosphatase (BAP) and osteocalcin values than subjects with a normal BMD. Biochemical assesment of bone metabolism may reveal useful in the early identification of coeliac patients who are at high risk of developing reduced BMD and bone impairment.

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