A B S T R A C T S   N U M E R O   7 - 8 / 1998

Diagnosis of viral hepatitis C and B infection by PCR Versus seromarkers in some chronic liver diseases

Mohamed Talaat Abdel Aziz, Randa Abdel kader Mahmoud, Magdy Amin El Serafy, Eman Mohamed Obayah
Medical Biochemistry, Cairo University

Biochimica Clinica 1998; 22(7-8): 413-421 [Article in english]

ABSTRACT. Correlation between the results of HBV and HCV seromarkers with their corresponding PCR results and the degree of association between HBV and /or HCV infection in hepatic schistosomiasis,liver cirrhosis with or without chronic active hepatitis and in mixed cases (bilharziasis+cirrhosis) were investigated. Sixty patients (52 males and 8 females) aged between 30 and 60 years suffering from chronic liver diseases were subjected to clinical, biochemical and ultrasonographic evaluations liver biopsy was done when possible.They were classified to 48 cirrhotic, 7 bilharzial and 5(mixed) patients. For all these subjects HBsAg, HBsAb, HBcAb, HBeAg, HBeAb by (ELISA) and anti-HCV by (ELISA2), HBV-DNA by PCR, HCV-RNA by RT-PCR were performed. We found among the 60 patients: 13 (21.67%) had HBV-DNA; 15 (25%) had HCV-RNA;13 (21.67%) had HBV and HCV and the last 19 (31.66%) had neither HBV nor HCV. Anti-HCV was positive in 13 patients with HCV-RNA negative and negative in 6 patients with HCV-RNA positive. We concluded that HBV markers were present in various combination with acceptable specificity and sensitivity especially HBsAg and HBeAg, yet PCR have increasingly used to assess HBV replication in patients infected with HBV mutant and also to detect trace amount of HBV-DNA.The diagnosis of HCV is not possible if serologically based, also, PCR is useful in detection of infection before the production of anti-HCV and in patients with persisting or fluctuating antibodies.Viral interference is common phenomenon in dual HBV and HCV infection and HCV seems to suppress HBV replication and induce seroclearance of its antigens. Cirrhosis is equally present in HBV and/or HCV positive patients by PCR. HCV infection is apparently more common in bilharzial patients compared to HBV infection.

Utility of anti-gliadin and anti-endomysium antibody tests for diagnosis of coeliac disease in a pediatric population

I. Barghini*, M. Niccolai, D. Monti, R. Rossetti
*U. O. Microbiologia - Spedali Riuniti - Azienda n. 3 - Piazza Giovanni XXIII - Pistoia

Biochimica Clinica 1998; 22(7-8): 422-424 [Article in italian]

ABSTRACT. Two hundred-sixty-nine children admitted at the Paediatric Unit of the Hospital of Pistoia were evaluated for a correlation between the clinical state and the serum levels of two coeliac disease markers, IgA and IgG AGA antibodies and IgA EMA antibodies. The AGA test was highly sensitive while the EMA test was the most specific for diagnosis of coeliac disease. In our experience AGA test may represent mainly a useful screening and monitoring method for coeliac disease, while EMA test must be used as a confirmation method.

Evaluation of the Eurogenetics Tosoh HL-723 GHb V A1C 2.2 system for the determination of glycated hemoglobin.

F. Caricato*, A. Bertolotto, L. Benzi, C. Mammini, M. Aragona, G. Penno, R. Navalesi
*Cattedra di Malattie del Metabolismo - Università di Pisa - U.O. Malattie del Ricambio e Gastroenterologia - Centro Regionale di Riferimento per il diabete mellito in età adulta - Azienda Ospedaliera Pisana

Biochimica Clinica 1998; 22(7-8): 425-430 [Article in italian]

ABSTRACT. Using 856 specimens obtained from patients sent to our Diabetes Service, we compared the concordance between the Diamat™ (Bio-Rad Laboratories), an automated analyzer measuring HbA1c by cation-exchange chromatography and the HLC-73 GHb V Tosoh (Eurogenetics), and HPLC method measuring HbA1c by cation-exchange cromatography on a non porous ion exchange column (TSK gel GLYCO Hsi) that reduces the time assay and ensures higher resolution of the HbA1c. Within- and between-batch CVs were <2% for the HLC-73 GHb V Tosoh (Tab. 1, 2). The regression equation and the correlation between the two systems resulted y=0.909x+0.158; r=0.99 respectively, but the regression line and the identity line did not coincide. Results obtained on the HLC-73 GHb V Tosoh were found to be negatively biased when compared with the Diamat™. The two methods when compared by Altman and Bland plots showed a mean bias = 0.466 (range:0.054-0.986) of HbA1c. Our results indicate that the two systems measure different proportion of HbA1c and, the lack of standardization of glyco-hemoglobin measurements remains a source of inter-methods variation.

Recommended IFCC-SIBioC methods for the measurement of catalytic activities of aminotransferases: implementation on the automatic Mega® Merck analyzer

R. Bonora*, M. Morandi, F. Pagani, M. Panteghini
*I Laboratorio Analisi Chimico-Cliniche, Spedali Civili, Brescia

Biochimica Clinica 1998; 22(7-8): 431-438 [Article in italian]

ABSTRACT. The aim of this study was to verify the possibility, using a fully automatic instrumentation, to get results strictly comparable to those obtained with IFCC recommended methods for aminotransferase (AST and ALT) determination, performed manually. After a preliminary evaluation of IFCC manual methods, implemented as reference, with good results of imprecision (CV < 1.7%) and bias (< 2%), we evaluated the proposed automated procedures. The linearity of the analytical respons e was good, with r > 0.9999, as well as the imprecision (between-day CV < 2.6% for AST and < 2.0% for ALT). The comparison with IFCC reference methods, according to NCCLS-EP9 protocol, using 40 samples with different activities, showed very good correlations (y = 1.004x - 0.26, r = 0.9998, for AST; y = 1.021x - 1.61, r = 0.9993, for ALT, respectively). The predicted bias was significantly lower than previously established performance criteria (SIBioC recommendation, æ 5% of IFCC method values) when evaluated at three different medical decision levels (-0.1 U/L and -0.6 U/L vs ± 2.5 U/L at 50 U/L, 0.1 U/L and 0.5 U/L vs ± 5 U/L at 100 U/L, 0.3 U/L and 1.5 U/L vs ± 7.5 U/L at 150 U/L, for AST and ALT, respectively). In conclusion, our data demonstrate the feasibility of the transfer of accuracy of IFCC methods into routine automatic procedures, through a careful work of instrumental adaptation of recommended methods.

Glycated hemoglobin determination

P. Metus*, P. Bonvicini, S. Guglielmo, M. Plebani
*Servizio di Medicina di Laboratorio - Azienda Ospedaliera - 35128 Padova

Biochimica Clinica 1998; 22(7-8): 439-442 [Article in italian]

ABSTRACT. We investigated the performance of a cation-exchange high performance liquid chromatographic system, (Automated A_1c 2.2-Eurogenetics-Tosoh) for HbA1c determination. The within assay (n = 30) and between assay (n = 20) coefficients of variation ranged from 0.9% to 1.2% and from 1.4% to 2.0% respectively. The assay was linear from 3.2 to 16.0% of HbA_1c. The effect of fetal hemoglobin was negligible at HbF concentration 30%. No effect of labile HbA_1c was found. The results from the evaluated system (y) were compared with those of the Bio-Rad. Diamat system (x) on fresh patiat samples. Linear regression analysis gave: y = 0.956x - 0.51y; r = 0.989; n = 95. Mean (±2SD) difference (x - y) was 0.88 (±0.48). The presence of abnormal hemoglobins as HbS, HbC, HbE and Hb Lepore was clearly revealed. In conclusion the Automated A_1c 2.2 system is suitable for chromatographic routine determination of glycated hemoglobin.

Autoimmune antibodies: frequency and patterns in an ederly population of both sexes

R. Rosso*, P. Cogorno, M.A. Pronzato*
*Laboratorio analisi chimico-cliniche e microbiologiche, Istituto Emanuele Brignole, P.le Brignole 2, 16125 Genova*Istituto di Patologia Generale, via L.B. Alberti 2, 16140 Genova

Biochimica Clinica 1998; 22(7-8): 443-449 [Article in italian]

ABSTRACT. The aim of our research was to check the frequency of autoantibody positivity in a sample-goup of randomly selected elderly subjects, By means of indirect fluorescence we tested for ANA, AMA, ASMA, n-DNA, and APCA antibodies a group of 100 blood samples from individuals of both sexes, of age ranging from 70 to 100 years, without any sign of autoimmune pathology. Overall positivity rate was 23% (23 subjects). The Western immunoblot technique was applied to the positive samples, in order to identify specific ENA autoantibodies. The results are discussed in relation to the different fluorescence patterns, and to the ENA detected. The utility of testing ederly peoples without clinical sign of autoimmune pathology for the presence of autoantibodies subjects is also discussed.

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