- Email: [email protected]
The new fully automated immunoassays enzymun-test® anti-HBs and HBsAg for ES systems
$128
P R E V A L E N T R I S K F A C T O R S IN H C V T R A N S M I S S I O N A M O N G A S E L E C T E D P O P U L A T I O N S A M P L E IN R O M E
G.Visco, M.A.Lonqo, P.Noto, G.Tossini, F.Ferri, U.Visco Comandini*, M.Zaccarelli "L.Spallanzani" Hosp.and Infect. Dis. Inst., "La Sapienza" University*, Rome, Italy. In 1989, whilst carrying out an epidemiological survey aimed at studying the seroprevalence for some viral antibodies among Italian Navy personnel, we found a low p r e v a l e n c e of HCVAb positivity (0,2 %) among 9,037 recruits, aged between 17-20 years. In this study, aimed to better understand the significance of some Risk Factors (R.F.) in HCV infection, an epidemiological transversal case-control survey was performed, through a questionnaire completed by 5,018 subjects presenting as outpatients to 5 Hospitals in Rome, tested for hepatitis viruses. The analysis of prevalences of the main R.F. between 152 subjects HCVAb+ve and 152 n e g a t i v e pair-matc.hed controls, shows a high prevalence of I.V.D.A., but only at younger ages. Within t~e non-IVDA sample, I.V. treatments with glass syringes (~<0.001), hospitalization (~<0.02), diabetes (~<0.003) (all before 1975) and transfusions before 1990(~<0.002) were the most significant R.F. in HCV infection. The role of dental cares, sexual partnership and cohabitation with HCVAb+ve persons did'nt appair from this stUdy.
INCIDENCE AND CLINICAL IMPACT OF HEPATITIS C FOLLOWING CADAVERIC KIDNEY TRANSPLANTATION: A SINGLE CENTER EXPERIENCE
G.H. Wimsber~er. J.M. Roob. J. Zeliko*. L. Kenner*. G. Hrfler*. Y, l~lShabrawi*. H, I-loiter. Deparunent of Internal Medicine and *Institute of F'athology Karl-Franzens-University. Graz, Austria. In renal transplantation chronic liver disease is associafed with increased morbidity and mortality. Since 1992. we longitudinally studied 408 patients from 7 Styrian hemodialysis units to determine the prevalence of hepatitis C virus (HCV) infection. Serum samples were tested for ,'mtibody to HCV by 2nd generation enzyme immunoassay (EIA lh Abbott) and for HCV-RNA by a two stage nested primer polymer,'tse chain reaction (PCR) assay giving an overall positivity rate of 18.9 ~.. Despite this high prevalence, liver function tests were Ilormal for prolonged periods in more than 95 ~ of PCR and/or EIA 11 positive subjects. Of these patients, 55 (17 females, 38 males) with persistently norm,'d serum transaminase levels received a cadaveric kidney (follo,,v-up 3 to 18 months). The basic immunosuppressive therapy consisted of methylprednisolone and Cyclosporine A. At the time of Iransplanwtion HCV-RNA was found positive in 5 patients, whereas HCV antitn~tlies were detected in 4 patients. Among the IICV-RNA positive uansplant recipients, marked elevated liver enzymes were seen in 3 cases within 1 to 4 months after transplantation (serum ALT above twice the upper limil of normal). Other causes for liver impairment such as CMV infection or drug induced hepatoxicity could be excluded. Two of 46 primarily I ICV ,tegative patients received a kidney from HCV-seroposilive donors, resulted in repeatedly positive testing for HCV-RNA sequences without seroconversion and detectable hepatic dysfunction. Our preliminary data suggest that the clinical course of IICV infection in immunosuppressed U'~utsplautrecipients is diffcreut from that of hemodialysis patients. Obviously HCV-PCR positive renal transplant recipients are at greater risk for posttransph'mt symptomatic bcpalifis duc to (re)activation of HCV infection. The long-term consequences of our findings e.g. interferon therapy prior to renal transplantation remain to be determined.
Papers read by title
QUANTITATIVE PCR AND GENOTYPE ANALYSIS DURING INTERFERON (ROFERON-2a) THERAPY OF CHRONIC HEPATITIS C INFECTION L. Wolfe, S. Lombardi, K. Gutekunst, with the HCV Diaenosfic Group 1, and I.C. Rvff2 with the Viral Hel~atitis Study Group 1Roche Molecular Systems, Somerville, New Jersey, USA and 2Hoffmann-La Roche, Basel, Switzerland In this study, we combined retrospective quantitative PCR and genotype analysis to provide additional tools for monitoring of patients treated with recombinant interferon alfa-2a (Roferon-A). From two multi-center treatment trials, serial samples from 265 patients were selected for analysis. Quantitation of serum HCV RNA was performed with a prototype assay utilizing a single primer pair, single enzyme (rTth), single amplification approach that incorporates an internal quantitation standard (IQS) into each amplification reaction, and a microweU plate detection format, using probes specific for HCV and for the IQS. HCV genotypes were assigned (based on 5'UTR sequence analysis) to 244 patients at baseline and, wherever possible, 3 months after therapy, with the following distribution of genotypes for this study-- la (17.6%), lb (58.6%), 2a (8.6%), 2b (0.8%), 3a (12.7%), 4 (1%), and 6 (0.4%).
THE NEW FULLY AUTOMATED IMMUNOASSAYS E N Z Y M U N - T E S T ® ANTI-HBs And HBsAg For ES Systems D Zdunek S Kohler. R Wehner. W Melchior R 8ablel Boehrmger Mannheim GmbH. Forschunszentrum Penzberg, FRG The new anti-HBs assay is a classical 1-step sandwich assay using streptav,din- coaled tube technology based on the use ol biolinylated and HRP-conlugaled h-HBsAg Ior the quanlltative determination of anlibod=es to Hepal~t,s 8 sudace antigen (HBsAg) in vitro The evaluation of the anti-HBs assay was carried out in 5 external laboratones and documented a good companbd,ly Io other enzyme ~mmunoassays (EIA) as well as RIA's The assay is working well Ior bolh. follow up of pat=ents wah acute and chrome Hepata,s B and vaccine reclp=ents showing comparable results with other routine methods. Specdlctty studies among blood donors offer a correlation of 99.5% with routine methods and a speoficity of > 99.5 %. The measuring range is extended trom 0 to approx. 400 IU/L with a analytical sensilivity of < 3 IU/L. The standards are cahbraled on the reference malenal WHO 1st IRP code 1042 Calibration is performed by a 2-point cahbrat=on curve based on hnear regress=on w=thoul recalibrat=on Applicat=ons are avadable lot a measunng temperature of 25"C and 37"C w,th a reduced mcubal=on time of 90 rain. The Enzymun-Test® HBsAg 2 Gen. is a qualitatfve 1-step sandwich immunoassay also based on streptavidin-coated tube technology wrlh biotmylated and HRP-conlugated antibodies for the determination ot Hepat,is B surface antigen. The new test shows a slgnihcant improvement in sensitivity and specillcity compared to the Enzymun-Tesf (=~HBsAg 1 Gen For determination ol spemhc,y 5400 sera were lested in 5 external laboratones Specificity of > 99.9% with unselecled blood donors and > 99.7% w=th hospitalised pal=ents was tound. The sensmvity was detected using seroconversion panels and dilution series of ddterent subtypes. The lower detection limit was lound at 0.1 E/rnl (PEI) which is comparable to other competitors. The controls are calibrated on the relerence material of the Paul Ehdich Institut A calculation of WHO units based on a conversion factor is available Ouantilication ol results is possible using a dilution series of the positive control as standards Conclusion' The new fully automated immunoassays antl-HBs and HSsAg show a great improvement concerning precision, sensitivity and specficity
P R E V A L E N T R I S K F A C T O R S IN H C V T R A N S M I S S I O N A M O N G A S E L E C T E D P O P U L A T I O N S A M P L E IN R O M E
G.Visco, M.A.Lonqo, P.Noto, G.Tossini, F.Ferri, U.Visco Comandini*, M.Zaccarelli "L.Spallanzani" Hosp.and Infect. Dis. Inst., "La Sapienza" University*, Rome, Italy. In 1989, whilst carrying out an epidemiological survey aimed at studying the seroprevalence for some viral antibodies among Italian Navy personnel, we found a low p r e v a l e n c e of HCVAb positivity (0,2 %) among 9,037 recruits, aged between 17-20 years. In this study, aimed to better understand the significance of some Risk Factors (R.F.) in HCV infection, an epidemiological transversal case-control survey was performed, through a questionnaire completed by 5,018 subjects presenting as outpatients to 5 Hospitals in Rome, tested for hepatitis viruses. The analysis of prevalences of the main R.F. between 152 subjects HCVAb+ve and 152 n e g a t i v e pair-matc.hed controls, shows a high prevalence of I.V.D.A., but only at younger ages. Within t~e non-IVDA sample, I.V. treatments with glass syringes (~<0.001), hospitalization (~<0.02), diabetes (~<0.003) (all before 1975) and transfusions before 1990(~<0.002) were the most significant R.F. in HCV infection. The role of dental cares, sexual partnership and cohabitation with HCVAb+ve persons did'nt appair from this stUdy.
INCIDENCE AND CLINICAL IMPACT OF HEPATITIS C FOLLOWING CADAVERIC KIDNEY TRANSPLANTATION: A SINGLE CENTER EXPERIENCE
G.H. Wimsber~er. J.M. Roob. J. Zeliko*. L. Kenner*. G. Hrfler*. Y, l~lShabrawi*. H, I-loiter. Deparunent of Internal Medicine and *Institute of F'athology Karl-Franzens-University. Graz, Austria. In renal transplantation chronic liver disease is associafed with increased morbidity and mortality. Since 1992. we longitudinally studied 408 patients from 7 Styrian hemodialysis units to determine the prevalence of hepatitis C virus (HCV) infection. Serum samples were tested for ,'mtibody to HCV by 2nd generation enzyme immunoassay (EIA lh Abbott) and for HCV-RNA by a two stage nested primer polymer,'tse chain reaction (PCR) assay giving an overall positivity rate of 18.9 ~.. Despite this high prevalence, liver function tests were Ilormal for prolonged periods in more than 95 ~ of PCR and/or EIA 11 positive subjects. Of these patients, 55 (17 females, 38 males) with persistently norm,'d serum transaminase levels received a cadaveric kidney (follo,,v-up 3 to 18 months). The basic immunosuppressive therapy consisted of methylprednisolone and Cyclosporine A. At the time of Iransplanwtion HCV-RNA was found positive in 5 patients, whereas HCV antitn~tlies were detected in 4 patients. Among the IICV-RNA positive uansplant recipients, marked elevated liver enzymes were seen in 3 cases within 1 to 4 months after transplantation (serum ALT above twice the upper limil of normal). Other causes for liver impairment such as CMV infection or drug induced hepatoxicity could be excluded. Two of 46 primarily I ICV ,tegative patients received a kidney from HCV-seroposilive donors, resulted in repeatedly positive testing for HCV-RNA sequences without seroconversion and detectable hepatic dysfunction. Our preliminary data suggest that the clinical course of IICV infection in immunosuppressed U'~utsplautrecipients is diffcreut from that of hemodialysis patients. Obviously HCV-PCR positive renal transplant recipients are at greater risk for posttransph'mt symptomatic bcpalifis duc to (re)activation of HCV infection. The long-term consequences of our findings e.g. interferon therapy prior to renal transplantation remain to be determined.
Papers read by title
QUANTITATIVE PCR AND GENOTYPE ANALYSIS DURING INTERFERON (ROFERON-2a) THERAPY OF CHRONIC HEPATITIS C INFECTION L. Wolfe, S. Lombardi, K. Gutekunst, with the HCV Diaenosfic Group 1, and I.C. Rvff2 with the Viral Hel~atitis Study Group 1Roche Molecular Systems, Somerville, New Jersey, USA and 2Hoffmann-La Roche, Basel, Switzerland In this study, we combined retrospective quantitative PCR and genotype analysis to provide additional tools for monitoring of patients treated with recombinant interferon alfa-2a (Roferon-A). From two multi-center treatment trials, serial samples from 265 patients were selected for analysis. Quantitation of serum HCV RNA was performed with a prototype assay utilizing a single primer pair, single enzyme (rTth), single amplification approach that incorporates an internal quantitation standard (IQS) into each amplification reaction, and a microweU plate detection format, using probes specific for HCV and for the IQS. HCV genotypes were assigned (based on 5'UTR sequence analysis) to 244 patients at baseline and, wherever possible, 3 months after therapy, with the following distribution of genotypes for this study-- la (17.6%), lb (58.6%), 2a (8.6%), 2b (0.8%), 3a (12.7%), 4 (1%), and 6 (0.4%).
THE NEW FULLY AUTOMATED IMMUNOASSAYS E N Z Y M U N - T E S T ® ANTI-HBs And HBsAg For ES Systems D Zdunek S Kohler. R Wehner. W Melchior R 8ablel Boehrmger Mannheim GmbH. Forschunszentrum Penzberg, FRG The new anti-HBs assay is a classical 1-step sandwich assay using streptav,din- coaled tube technology based on the use ol biolinylated and HRP-conlugaled h-HBsAg Ior the quanlltative determination of anlibod=es to Hepal~t,s 8 sudace antigen (HBsAg) in vitro The evaluation of the anti-HBs assay was carried out in 5 external laboratones and documented a good companbd,ly Io other enzyme ~mmunoassays (EIA) as well as RIA's The assay is working well Ior bolh. follow up of pat=ents wah acute and chrome Hepata,s B and vaccine reclp=ents showing comparable results with other routine methods. Specdlctty studies among blood donors offer a correlation of 99.5% with routine methods and a speoficity of > 99.5 %. The measuring range is extended trom 0 to approx. 400 IU/L with a analytical sensilivity of < 3 IU/L. The standards are cahbraled on the reference malenal WHO 1st IRP code 1042 Calibration is performed by a 2-point cahbrat=on curve based on hnear regress=on w=thoul recalibrat=on Applicat=ons are avadable lot a measunng temperature of 25"C and 37"C w,th a reduced mcubal=on time of 90 rain. The Enzymun-Test® HBsAg 2 Gen. is a qualitatfve 1-step sandwich immunoassay also based on streptavidin-coated tube technology wrlh biotmylated and HRP-conlugated antibodies for the determination ot Hepat,is B surface antigen. The new test shows a slgnihcant improvement in sensitivity and specillcity compared to the Enzymun-Tesf (=~HBsAg 1 Gen For determination ol spemhc,y 5400 sera were lested in 5 external laboratones Specificity of > 99.9% with unselecled blood donors and > 99.7% w=th hospitalised pal=ents was tound. The sensmvity was detected using seroconversion panels and dilution series of ddterent subtypes. The lower detection limit was lound at 0.1 E/rnl (PEI) which is comparable to other competitors. The controls are calibrated on the relerence material of the Paul Ehdich Institut A calculation of WHO units based on a conversion factor is available Ouantilication ol results is possible using a dilution series of the positive control as standards Conclusion' The new fully automated immunoassays antl-HBs and HSsAg show a great improvement concerning precision, sensitivity and specficity