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An automated sample preparation method for HCV PCR
Virul
(
hepatitis:
co6/077
clinical
205
uspects
( CO6/079 1
1
IRON STATUS OF PATIENTS HEPATITIS C
AN AUTOMATED SAMPLE PREPARATION FOR HCV PCR
WITH CHRONIC
V.T. Ivashkin*. E. Lukina. M. Maievskaja*. E. Svsoieva. C. Pavlov*. N.D. Khoroshko.
A. Levina, G.A. Frank.
Moscow Medical Academy*, National Research Center for Haematology, Moscow, Russia. In spite of the ever-increasing body of knowledge on the subject of iron, infection and neoplasia, many aspects remain unclear and in some cases controversial. Recent studies suggest that increased hepatic iron may impair the response to interferon therapy in patients (pts) with chronic hepatitis C and there is a significant correlation between hepatic iron grade and serum ferritin. The goal of this study was to investigate the iron status of 24 untreated noncirrhotic pts with serum HCV RNA positive chronic hepatitis C. All the pts (age 23-67 years) had low histological activity index (Knodell score = 3-8) and serum aminotransferases
HCV INFECTION EPIDEMIOLOGICAL
METHOD
IN FIAEMODIALYSED AND CLINICAL ASPECTS
PATIENTS
-
M.Indot’ . MZalewska . U.Bem?. A.Gladvsz' .D.Zwoliliskaz Dpt.of Infectious Diseases’ , Dpt of Nephrolo&, Medical University, Wrociaw, Poland A’iM: HCV infection is an important problem in I-ID unitsThe aim of the study was to evaluate epidemiology and course of HCV infection in young, haemodialysed patients. MATERIAL AND METHODS: 18 patients, age range 9 - 41, x=17,9, M 8,F 10 were evaluated. Anti-HCV antibodies were tested by screening enzyme immunoassay ( Abbott Diagnostic HCV EIA, 3’d gen.). AntiHCV antibodies were sought by Liatek HCV 3” gen.( Organon Teknika). HCV RNA was detected by a standarized HCV RNA PCR assay ( Amplicor HCV, Roche Diagnostic System) Amplicor HCV Monitor (Troche Diagnostic System) was used to monitor viral load, the results were expressed in HCV RNA copies/ml. RESULTS: HCV infection was diagnosed in 11 ( 6 1 %) patients. , the average duration of haemodialysis in infected patients was 5 years, in HCV negative only 3 months.The other risk factors of HCV infection (blood transfusion,surgery, IVDU) were analysed - there was no correlations beteween HCV infection and these factors. In two cases HCV infection was diagnosed by PCR anti-HCV were negative. None of the HCV infected patients have the clinical manifestation of hepatitis, the mild activity of ALT was observed in 4 cases only. HCV viremia was significantly lower than in control group ( HCV infected. nonhaemodialysed patients ) 3 18,6 copies/ml YS 1011.2 copies/ml @
JR. Hildebrandt. *R. Herron, A. Starron. Z. Li, E. Reitz. T. Niemiec. M. Fairchild, K.C. Youna, G. Baechler. *A. Decker, *K. Loaan. l*W. Rev. Roche Molecular Systems, Alameda, California, USA; *Branchburg, NJ, USA; l * Roche Diagnostics Instrument Center Tegimenta, Rotkreuz, Switzerland An instrument is being developed to accomplish fully automated specimen processing for isolation of Hepatitis C Virus (HCV) RNA. The specimen preparation method is based on hybridization of viral target with oligonucleotide probe followed by magnetic particle capture of the probeRNA complex. The processed specimen is compatible with the AMPLICOR@ HCV, COBAS AMPLICORTM HCV, AMPLICOR HCV MONITORTM, and COBAS AMPLICOR HCV MONITORTM Tests. A patient correlation study showed good correlation between the automated method and the AMPLICOR HCV MONITORTM Test with manual specimerl preparation. Linearity and precision over the dynamic range of the AMPLICOR HCV MONITORTM Test will be presented. The automated specimen preparation method increased the sensitivity of the COBAS AMPLICORTM HCV Test to better than 100 copies/ml without detectable sample inhibition. Full automation of specimen preparation in conjunction with amplification and detection on the COBAS AMPLICORTM analyzer will insure further incorporation of PCR in the routine diagnostic laboratory.
Elevated Alanine Aminotransferase (ALT) in Blood Donors: An Assessment of the Main Causes and Its Relationship to the Development of Hepatitis. E L. Goncales Jr.. R. S. B. Snack. PM. 0. Pamrordanou. M. H. P. PavanM. C. Chmvm. C. A. F. Escanhmia, ‘N. S. L. Goncdes. “J. R. R. Pmho. Faculdade de Ci&ias Medicas, Univasidade Estadual de Campinas, S&o Paula Brazil, *Hemccmtm Univasidade Estadual de Camhas: **Serviw de Virolonia, lmtituto Adolf0 Lutz, Secretaria Estadul de Sairde, Sio Paula, SP, Ekil.
determination of aminotranferases levels is very useful in the dignoses of hepatopathies. In recent years, an elevated serum AL.T levels in blood donors has been associated with an increase risk of post-transfusion hepatitis The purpose of the study was to assesses the factors associated with elevated ALT levels in a cohort of voluntary blood donors and evaluates the relationship between rmsed ALT levels and the development of HCV infection. Materials and Methods: 116 volunteer blood donors with elevated ALT at the time of their first donation were studied. All of the donors were questioned about previous hepatopathies, exposure to hepatitis, exposure to chemicals, use of medication or drugs, sexual behaviour, contact with blood or secretions and their intake of alcohol. Every three months the serum levels of AST, XT, alkaline phosphatase, a glutanil transpeptidase, cholesterol, triglyceride and glycemia are assessed over a two years follow-up. The serum thyroid hormone levels as well as the presence of auto-antibodies were measured. Abdominal ultrasound was performed in patients with persistently elevated ALT or AST levels. A needle biopsy of liver using the Menghine technique was performed in nine individuals. Anti-HCV antibody levels were assayed at the first clinical evaluation and during the follow-up period (about six months later) Rerulrs: None of donors listed positive for hepatitis B and hepatits C markers during the follow-up. 10 1I1 16 (87%) continued to exhibit elevated ALT levels during the follow-up. Obesity and alcoholism were the principal factors related to elevated ALT levels in 91/101 (90%) of the donors. 3001 (2,9%) were exposed daily to hepatotoxic industrial solvents and paints. Hiperlipidemia (l/101), hypothyoidism (l/101), diabetes mellitus (l/101) also were associated with increased ALT levels. In 31101 (2,9%) the liver biopsy showed reactive hepatitis. Only one donor out of 116 (0.9%) had non A-G viral hepatitis and elevated ALT levels during the follow-up period Conclusion: The determination of aminotransferases levels isn’t helpful1 in preventing post-transfusion hepatitis and must be discontinued in Brazil. The
(
hepatitis:
co6/077
clinical
205
uspects
( CO6/079 1
1
IRON STATUS OF PATIENTS HEPATITIS C
AN AUTOMATED SAMPLE PREPARATION FOR HCV PCR
WITH CHRONIC
V.T. Ivashkin*. E. Lukina. M. Maievskaja*. E. Svsoieva. C. Pavlov*. N.D. Khoroshko.
A. Levina, G.A. Frank.
Moscow Medical Academy*, National Research Center for Haematology, Moscow, Russia. In spite of the ever-increasing body of knowledge on the subject of iron, infection and neoplasia, many aspects remain unclear and in some cases controversial. Recent studies suggest that increased hepatic iron may impair the response to interferon therapy in patients (pts) with chronic hepatitis C and there is a significant correlation between hepatic iron grade and serum ferritin. The goal of this study was to investigate the iron status of 24 untreated noncirrhotic pts with serum HCV RNA positive chronic hepatitis C. All the pts (age 23-67 years) had low histological activity index (Knodell score = 3-8) and serum aminotransferases
HCV INFECTION EPIDEMIOLOGICAL
METHOD
IN FIAEMODIALYSED AND CLINICAL ASPECTS
PATIENTS
-
M.Indot’ . MZalewska . U.Bem?. A.Gladvsz' .D.Zwoliliskaz Dpt.of Infectious Diseases’ , Dpt of Nephrolo&, Medical University, Wrociaw, Poland A’iM: HCV infection is an important problem in I-ID unitsThe aim of the study was to evaluate epidemiology and course of HCV infection in young, haemodialysed patients. MATERIAL AND METHODS: 18 patients, age range 9 - 41, x=17,9, M 8,F 10 were evaluated. Anti-HCV antibodies were tested by screening enzyme immunoassay ( Abbott Diagnostic HCV EIA, 3’d gen.). AntiHCV antibodies were sought by Liatek HCV 3” gen.( Organon Teknika). HCV RNA was detected by a standarized HCV RNA PCR assay ( Amplicor HCV, Roche Diagnostic System) Amplicor HCV Monitor (Troche Diagnostic System) was used to monitor viral load, the results were expressed in HCV RNA copies/ml. RESULTS: HCV infection was diagnosed in 11 ( 6 1 %) patients. , the average duration of haemodialysis in infected patients was 5 years, in HCV negative only 3 months.The other risk factors of HCV infection (blood transfusion,surgery, IVDU) were analysed - there was no correlations beteween HCV infection and these factors. In two cases HCV infection was diagnosed by PCR anti-HCV were negative. None of the HCV infected patients have the clinical manifestation of hepatitis, the mild activity of ALT was observed in 4 cases only. HCV viremia was significantly lower than in control group ( HCV infected. nonhaemodialysed patients ) 3 18,6 copies/ml YS 1011.2 copies/ml @
JR. Hildebrandt. *R. Herron, A. Starron. Z. Li, E. Reitz. T. Niemiec. M. Fairchild, K.C. Youna, G. Baechler. *A. Decker, *K. Loaan. l*W. Rev. Roche Molecular Systems, Alameda, California, USA; *Branchburg, NJ, USA; l * Roche Diagnostics Instrument Center Tegimenta, Rotkreuz, Switzerland An instrument is being developed to accomplish fully automated specimen processing for isolation of Hepatitis C Virus (HCV) RNA. The specimen preparation method is based on hybridization of viral target with oligonucleotide probe followed by magnetic particle capture of the probeRNA complex. The processed specimen is compatible with the AMPLICOR@ HCV, COBAS AMPLICORTM HCV, AMPLICOR HCV MONITORTM, and COBAS AMPLICOR HCV MONITORTM Tests. A patient correlation study showed good correlation between the automated method and the AMPLICOR HCV MONITORTM Test with manual specimerl preparation. Linearity and precision over the dynamic range of the AMPLICOR HCV MONITORTM Test will be presented. The automated specimen preparation method increased the sensitivity of the COBAS AMPLICORTM HCV Test to better than 100 copies/ml without detectable sample inhibition. Full automation of specimen preparation in conjunction with amplification and detection on the COBAS AMPLICORTM analyzer will insure further incorporation of PCR in the routine diagnostic laboratory.
Elevated Alanine Aminotransferase (ALT) in Blood Donors: An Assessment of the Main Causes and Its Relationship to the Development of Hepatitis. E L. Goncales Jr.. R. S. B. Snack. PM. 0. Pamrordanou. M. H. P. PavanM. C. Chmvm. C. A. F. Escanhmia, ‘N. S. L. Goncdes. “J. R. R. Pmho. Faculdade de Ci&ias Medicas, Univasidade Estadual de Campinas, S&o Paula Brazil, *Hemccmtm Univasidade Estadual de Camhas: **Serviw de Virolonia, lmtituto Adolf0 Lutz, Secretaria Estadul de Sairde, Sio Paula, SP, Ekil.
determination of aminotranferases levels is very useful in the dignoses of hepatopathies. In recent years, an elevated serum AL.T levels in blood donors has been associated with an increase risk of post-transfusion hepatitis The purpose of the study was to assesses the factors associated with elevated ALT levels in a cohort of voluntary blood donors and evaluates the relationship between rmsed ALT levels and the development of HCV infection. Materials and Methods: 116 volunteer blood donors with elevated ALT at the time of their first donation were studied. All of the donors were questioned about previous hepatopathies, exposure to hepatitis, exposure to chemicals, use of medication or drugs, sexual behaviour, contact with blood or secretions and their intake of alcohol. Every three months the serum levels of AST, XT, alkaline phosphatase, a glutanil transpeptidase, cholesterol, triglyceride and glycemia are assessed over a two years follow-up. The serum thyroid hormone levels as well as the presence of auto-antibodies were measured. Abdominal ultrasound was performed in patients with persistently elevated ALT or AST levels. A needle biopsy of liver using the Menghine technique was performed in nine individuals. Anti-HCV antibody levels were assayed at the first clinical evaluation and during the follow-up period (about six months later) Rerulrs: None of donors listed positive for hepatitis B and hepatits C markers during the follow-up. 10 1I1 16 (87%) continued to exhibit elevated ALT levels during the follow-up. Obesity and alcoholism were the principal factors related to elevated ALT levels in 91/101 (90%) of the donors. 3001 (2,9%) were exposed daily to hepatotoxic industrial solvents and paints. Hiperlipidemia (l/101), hypothyoidism (l/101), diabetes mellitus (l/101) also were associated with increased ALT levels. In 31101 (2,9%) the liver biopsy showed reactive hepatitis. Only one donor out of 116 (0.9%) had non A-G viral hepatitis and elevated ALT levels during the follow-up period Conclusion: The determination of aminotransferases levels isn’t helpful1 in preventing post-transfusion hepatitis and must be discontinued in Brazil. The