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Evaluation of mRNA for Prostate Specific Membrane Antigen (mRNA-PSM) assay in peripheral blood from patients with Prostate Carcinoma

Gabriella Priolo, Nicola Faraone, Giuseppe Aimo*, Carlo Terrone, Gianni Pelucelli, Teresa Zaccaria, Eleonora Ciocia, Salvatore Rocca Rossetti, Roberto Pagni
U.O.A. Laboratorio Centrale di Chimica Clinica "Baldi e Riberi"

Biochimica Clinica: 2000; 24(2): 77-82 [Article in italian]

ABSTRACT. In order to verify the clinical value of the mRNA-PSM detection with RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction) in the management of prostate carcinoma we performed the assay in peripheral blood samples from patients with different stage prostate cancer (48 cases) in comparison with patients affected by benign urological diseases (42 cases) and healthy volunteers (18 cases). We did not obtaine false positive results neither in healthy volunteers, males and females, nor in benign non prostatic diseases. Also in benign prostate hypertrophy (BPH) the specificity was very high (91.7%). The sensitivity of the assay was 100% both in metastatic cancer and in a group of clinically confined prostate cancer before radical prostatectomy. The assay was positive in 65.2% (15/23) of the patients after radical prostatectomy with no clinical evidence of recurrence at the moment of the blood sampling. According to the most diffused opinion in the literature our study did not demonstrate, although in a limited number of cases, any significant statistical correlation between the result of the test and the staging or the Gleason score of the cancer. In conclusion, the results don't allow to assign to the test a sufficient clinical value to introduce it in the decisional process for the management of the prostate carcinoma.

Measurement of 25-hydroxyvitamin D: comparison between the reference-RIA method and an HPLC method

Milena Romanello, Gianpietro Colautti, Luigi Moro^1,2
1Dipartimento di Biochimica, Biofisica e Chimica delle Macromolecole, Università degli Studi di Trieste, 34127 Trieste
²Centro per lo Studio delle Malattie Metaboliche dell'Osso, Università degli Studi di Trieste, Azienda per i Servizi Sanitari n. 2 "Isontina", 34170 Gorizia

Biochimica Clinica: 2000; 24(2): 83-86 [Article in italian]

ABSTRACT. The measurement of circulating 25-hydroxyvitamin D (25-OH Vit D) is used for diagnosis of hypovitaminosis D which constitutes a major risk factor for osteopenia and bone fracture. The development of an analytical method, accurate and easy to apply, should be useful to measure the vitamin D status in patients presenting the risk of osteoporosis. We have performed a study for the evaluation of an HPLC test for the 25-OH Vit D measurement. We have determined the following perfomance characteristics: within-run and between-run precision, linearity and detection limit. Moreover, we have compared the HPLC results with those obtained with a RIA method, which is the reference one. The HPLC test proves to be sensitive, accurate, linear to values of 25-OH Vit D so high to be toxic, and easy to perform. The correlation with the RIA method seems to be good even though not optimal, but this is overcome by the advantages of precision of the HPLC method, especially useful in the longitudinal monitoring of patients which are treated with vitaminic compounds.

Updates on Laboratory Medicine and Doping. The new CONI Campaign "Io non rischio la salute!"

Giuseppe Lippi* e Giancesare Guidi
*Istituto di Chimica e Microscopia Clinica, Dipartimento di Scienze Biomediche e Morfologiche, Università degli Studi di Verona, Verona

Biochimica Clinica: 2000; 24(2): 87-95 [Article in italian]

ABSTRACT. Few months afterward the worrying re-explosion of the "doping scandal", following the exclusion of the already winner of the 1999 edition of the Giro d'Italia, Marco Pantani, the approach to face the heterogeneous problem of doping in sport has changed. Part of these changes come from the numerous criticisms issued on the doubtful actions of the Antidoping Scientific Committee, who has sanctioned athletes on the only base of abnormal hematocrit values, as only proof of erythropoietin consumption or blood doping. The numerous reproaches, often authoritative and intended to abjure the inaccurate and non-rigorous approach, have produced some tangible effects. The notorious Campaign "Io non rischio la salute!" has shown some uncertainties, especially in terms and time of application and the formal accusations, supported by reliable scientific bases, have forced CONI to a substantial revision of the protocol. These changes concerned also Laboratory Medicine that, as expected, is now "reassuming its prominent role", as attested by the recent inclusion of influential representatives of this discipline in the Scientific Committee and by the more prominent role occupied by the Laboratory Medicine in the revised version of the protocol. The main aspects of the new Champaign are synthesized and discussed in the following parts of this article.

An external quality assessment program for DNA analysis with PCR amplification

Claudio Orlando, Claudia Casini Raggi, Mario Pazzagli*
*Unità di Biochimica Clinica, Dipartimento di Fisiopatologia Clinica, Università di Firenze

Biochimica Clinica: 2000; 24(2): 96-104 [Article in italian]

ABSTRACT. In contrast to the large variety of possible applications of polymerase chain reaction (PCR) technique in diagnostic laboratories, external quality assessment (EQA) programs are until now limited to only few tests with established clinical value. Considering this restriction, EQA programs should be directed towards the evaluation of analytical aspects which are common to the great majority of tests based on PCR. We have developed an EQA program for the evaluation of DNA extraction, its amplification and the analysis of products after PCR. The program presents three control levels: 1. DNA extraction (quality and quantity); 2. PCR performances (specificity and efficiency); 3. interpretation of the results after electrophoresis. The first lot of reagents was sent to 35 Italian laboratories. Of these, 16 have completed the tests required. The analysis of the data collected shows a fair homogeneity with regard to DNA extraction, but a great variability in efficiency and specificity of the amplification procedures and results analysis. These preliminary results seem to confirm the validity of the approach of this EQA program.

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