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Detection of hepatitis C virus using an automated PCR instrument system
512A
1621
AASLD
ABSTRACTS
A RANDOMIZED STUDY COMPARING A FIXED-DOSE OF RECOMBINANT INTERFERON ALPHA 2a (ROFERON) VERSUS A DECREASING DOSE SCHEDULE IN PATIENTS WITH CHRONIC HEPATITIS C : A MULTICENTER STUDY IN CHINA. GB YAOt. DZ. XU2. J. GAO3. LM. ZHANG1. XH. WU4, OB. ZHANGd AND DC. HU5 qingan Central Hosptial, 2Ditan Hosptial, 3Ruijin Hospital, 4Huashan Hospital, SZhongshan Hospital.
1622
Conclusion : Recombinant interferon alpha 2a (Roferou) is effective in the treatment of chronic hepatitis C, the recommended dosage in most Chinese patients will be 3 MU t.i.w, for 6 months, But in some poor response patients, escalating dosage and prolonged schedule will be needed.
HMG-CoA REDUCTASE INHIBI.TOR (SIMVASTATIN) IMPROVED ALCOHOLIC LIVER DISEASE. T. Yuki, H. A d a c h i , T. Y o s h i z a w a l l T. K a w a ~ u c h i 2 1 T . O ~ i h a r a . A. M i v a z a k i . S. W a t a n a h e a n d N. Sato3~ 1 ) M e d i c i n e a n d 2 ) R a d i o l o g y , K a t s u n a n M u n i c i p a l H o s p i t a l , Chlba, and 3 ) D e p t . o f G a s t r o e n t e r o l o g y , J u n t e n d 0 U n i v e r s i t y , S c h o o l o f M e d i c i n e , Tokyo, J a p a n AIM: HMG-CoA r e d u c t a s e i n h i b i t o r i s c l i n i c a l l y u s e d f o r t h e t r e a t m e n t o f h y p e r l i p i d e m i a a n d w e l l known t o i n h i b i t cholesterol s y n t h e s i s and s c a v e n g e f r e e r a d i c a l s . On t h e other hand, alcoholic fatty liver often accompanies h y p e r l i p l d e m i a a n d l i p i d p e r o x i d a t i o n due t o f r e e r a d i c a l i s one o f mechanisms o f a l c o h o l i c l i v e r d i s e a s e , e s p e c i a l l y alcoholic hepatitis. We t r i e d t o c l a r l f y t h e e f f e c t o f S i m v a s t a t i n on p a t i e n t s with alcoholic fatty liver and hepatitis. S u b j e c t s a n d Methods: 22 o u t p a t i e n t s , who were d i a g n o s e d a s f a t t y l i v e r and h e p a t i t i s u l t r a s o n o g r a p h i c a l l y and liver biopsy respectively. G r o u p - l : 16 p a t i e n t s w i t h a l c o h o l i c a n d n e g a t i v e HCV-Ab (2nd g e n e r a t i o n ) . Group-2: 5 p a t i e n t s w l t h a l c o h o l i c a n d p o s i t i v e HCV-Ab. Group-3: 1 p a t i e n t w i t h a l c o h o l i c h e p a t i t i s a n d HCV-Ab. S i m v a s t a t l n ( 5 - 1 0 m g / d a y ) was g i v e n t o p a t i e n t s o f G r - 1 a n d Gr-2 f o r 4 months and that of Gr-3 for 1 year and 8 months. R e s u l t s : 1) I n b o t h Gr-1 and G r - 2 , serum T.G. LDL, VLDL, AST, ALT, G-GTF, a s w e l l a s l l p i d p e r o x i d e were h i g h e r t h a n normal level. Simvastatin treatment could lower those c h a n g e s b y 10-30%. 2) Hepatic CT v a l u e a t S 7 was r e c o v e r e d hy Simvastatin in Gr-1. (initial vales vs treated value: 5 5 . 0 ± 3 . 6 HU a n d 6 5 . 7 ~ 2 . 4 HU, p < 0 . 0 5 , r e s p e c t i v e l y ) 3) In Gr-3, Simvasta%in could improve liver pathology ( s w e l l i n g a n d n e c r o s i s o f h e p a t o e y t e s , PMN a n d l y m p h o c y t e s inf$1tration) and hyperlipidemia. C o n c l u s i o n : These d a t a indicate continuously drinking inducing alcoholic fatty liver and hepatitis with hyperlipidemia and lipidperoxidation, may lead to liver cell injury. S i m v a s t a t i n improved t h o s e c h a n g e s i n a l c o h o l i c f a t t y l i v e r a n d h e p a t i t i s w i t h a n d w i t h o u t HCV i n f e c t i o n .
DETECTION OF HEPATITIS C VHiUS USING AN AUTOMATED PCR INSTRUMENT SYSTEM. C. Young. S. Barmat, T. Haley, S. Lombardi. and L. Wolfe. Roche Molecular Systems, Inc., Branchburg, New Jersey, 08876-1760 Three hundred thirty eight serum or plasma specimens were tested for the presence of Hepatitis C Vires (HCV) using a prototype HCV Polymerase Chain Reaction (PCR) assay on an automated instrument system (COBAS® AMPLICORTM). The COBAS AMPLICOR Instrument System contains two independently controlled thermal eyelet blocks, a robotic arm for reagent handling, and a flexible analyzer for detection, thereby, integrating amplification, detection, and data/reagent management into an automated format that provides "walk away" capability to the user. The prototype HCV PCR assay was a coamplification of HCV and an internal control (IC) designed to monitor the amplification capability of each processed specimen. Sensitivity testing using RNA transcript demonstrated that the system was capable of detecting 5 input copies ofHCV RNA 100% of the time and inchisivity studies have shown that the assay detects all common HCV genotypes. The clinical specimens were evaluated by the automated prototype HCV assay in a blind study and the results were compared to the AMPLICORT M HCV Test in the mierowell plate format. When available, EL& and RIBA testing were used to resolve discrepant results bctween the two tests. Ofthe 222 AMPLICOR HCV negative samples tested, there was one false positive result using the prototype HCV test, for a specificity of 99:5% for this sample population: All 116 HCV positive samples were positive using the prototype HCV test, for a sensitivity of 100% enmpared to the AMPLICOR HCV Test. Overall, there was a 98.8% concordance between the AMPLICOR HCV Test and the prototype automated assay during initial testing and 100% concordance upon repeat testing. Ofthe 338 specimens tested, 3 had low IC values requiring repeat extraction and testing. Upon repeat, all 3 IC values recovered to normal levels. None of the 338 clinical specimens were determined to be PCR inhibited based on IC results. Overall, the prototype PCR assay on the COBAS AMPLICOR Instrument System performed equivalently to the AMPLICOR HCV Test in the microwell plate format for the deteetien of HCV in human serum or plasma.
It has been proven that interferon alpha 2a is effective in the treatment of chronic hepatitis C(HC), however the appropriate dosage and sehednle in Chinese patients remaines to be determined. The object of this study was to compare the efficacy of a dose of 3 MU of Roferon three times a week for 6 months with a starting dose of 6 MU for 3 months and a subsequent reduction to 3 MU three times a week for further 3 months. Sixty-eight patients with serological and histological proof of chronic hepatitis C with elevated serum ALl" levels were randomized into tWOgroups. 63 patients completed the full course of treatment. 5 patients were withdrawn from the study, 4 due to personal reasons and one due to aggravation of disease during treatment. 34 patients received 3 MU Referen t.i.w, for 6 months (group A), and 29 patients received 6 MU Roferon t.i.w. 3 months followed by 3 MU t.i.w, for another 3 months (group B). The sex, age, serum bilirubin, ALT and AST levels were matched in both groups. At the end of the 6th month the complete response and: partial response rates in group A were 67.6% and 8.8% respectively, the clearance of serum HCV-RNA was 71.4%. In group B, the complete and partial response rates were 62% and 20.7% and the elearanee of HCV-RNA was 72%. The efficacy between two dosage groups showed no statistical difference. The common adverse reactions (ADRS) were also similar in both groups. Most ADRS were tolerable only one patient suffered severe leucopenia requiringdose reduction. Another one patient suffered from marked elevation of serum bilirobin and ALT and aggravation of clinical symptoms, who was withdrawn from treatment. The response rates of Roferon treatment were correlated with genotype of HCV, severity of histopatho!ogy and titer of antibiodies to interferon.
1623
HEPATOLOGY O c t o b e r 1 9 9 5
1624
QUANTITATION OF ANTI-CORE HCV ANTIBODIES IS USEFUL FOR MONITORING IFN-THERAPY IN C H R O N I C HEPATITIS C. R Zachoval, R Hoffmann, G Michel, W Swoboda, H Dieoolder. MC Juno. G Frrsner and G R Pape. Medizinische Klinik II, Klinikum G-roBhadern and M a x von Pettenkofer-Institut, University o f Munich; Abbott Laboratories, Wiesbaden; Germany. Interferon.alpha (IFN) is an effective treatment in 20-30% o f patients with chronic hepatitis C (cHC). Evaluation o f effectiveness is based on serial alanine aminotransferase (ALT) measurements and determination o f H C V R N A in the serum. Serological markers o f efficacy are badly needed because they better reflect the patient's immune response to H C V and therapy. We therefore evaluated the usefulness o f quantitating antibodies to H C V core o f the I g G and separately o f the I g M type (automated immunoassays on the IMx analyser, Abbott Laboratories) during the course o f IFN therapy and up to at least 6 months after t h e end o f treatment. Thirty-six patients were included in the study: 10/36 were sustained responders (SR) to a course o f 3-5 Mio IFN s.c., thrice weekly for 6-12 months whereas 14 patients were classified as transient responders (TR) and 12 individuals as nonresponders (NR). The titer o f anti-HCV core IgM and I g G declined significantly (p<0.05, MarmWhitney U-test) at the end o f treatment in TR and SR compared to NR. Clinical relapse in TR was associated with an early increase o f both types o f specific antibodies whereas they remained at a constant low level in SR. In our study population we were not able to demonstrate a correlation between the total amount o f antibodies against H C V core before treatment and sustained response or the viral load in the serum (Amplicor Monitor, Hoffmarm-La Roche, Switzerland). Our data demonstrate that sequential quantitative assessment o f antiH C V core I g G and IgM is a valuable tool in monitoring responsiveness to IFN therapy in eriC.
1621
AASLD
ABSTRACTS
A RANDOMIZED STUDY COMPARING A FIXED-DOSE OF RECOMBINANT INTERFERON ALPHA 2a (ROFERON) VERSUS A DECREASING DOSE SCHEDULE IN PATIENTS WITH CHRONIC HEPATITIS C : A MULTICENTER STUDY IN CHINA. GB YAOt. DZ. XU2. J. GAO3. LM. ZHANG1. XH. WU4, OB. ZHANGd AND DC. HU5 qingan Central Hosptial, 2Ditan Hosptial, 3Ruijin Hospital, 4Huashan Hospital, SZhongshan Hospital.
1622
Conclusion : Recombinant interferon alpha 2a (Roferou) is effective in the treatment of chronic hepatitis C, the recommended dosage in most Chinese patients will be 3 MU t.i.w, for 6 months, But in some poor response patients, escalating dosage and prolonged schedule will be needed.
HMG-CoA REDUCTASE INHIBI.TOR (SIMVASTATIN) IMPROVED ALCOHOLIC LIVER DISEASE. T. Yuki, H. A d a c h i , T. Y o s h i z a w a l l T. K a w a ~ u c h i 2 1 T . O ~ i h a r a . A. M i v a z a k i . S. W a t a n a h e a n d N. Sato3~ 1 ) M e d i c i n e a n d 2 ) R a d i o l o g y , K a t s u n a n M u n i c i p a l H o s p i t a l , Chlba, and 3 ) D e p t . o f G a s t r o e n t e r o l o g y , J u n t e n d 0 U n i v e r s i t y , S c h o o l o f M e d i c i n e , Tokyo, J a p a n AIM: HMG-CoA r e d u c t a s e i n h i b i t o r i s c l i n i c a l l y u s e d f o r t h e t r e a t m e n t o f h y p e r l i p i d e m i a a n d w e l l known t o i n h i b i t cholesterol s y n t h e s i s and s c a v e n g e f r e e r a d i c a l s . On t h e other hand, alcoholic fatty liver often accompanies h y p e r l i p l d e m i a a n d l i p i d p e r o x i d a t i o n due t o f r e e r a d i c a l i s one o f mechanisms o f a l c o h o l i c l i v e r d i s e a s e , e s p e c i a l l y alcoholic hepatitis. We t r i e d t o c l a r l f y t h e e f f e c t o f S i m v a s t a t i n on p a t i e n t s with alcoholic fatty liver and hepatitis. S u b j e c t s a n d Methods: 22 o u t p a t i e n t s , who were d i a g n o s e d a s f a t t y l i v e r and h e p a t i t i s u l t r a s o n o g r a p h i c a l l y and liver biopsy respectively. G r o u p - l : 16 p a t i e n t s w i t h a l c o h o l i c a n d n e g a t i v e HCV-Ab (2nd g e n e r a t i o n ) . Group-2: 5 p a t i e n t s w l t h a l c o h o l i c a n d p o s i t i v e HCV-Ab. Group-3: 1 p a t i e n t w i t h a l c o h o l i c h e p a t i t i s a n d HCV-Ab. S i m v a s t a t l n ( 5 - 1 0 m g / d a y ) was g i v e n t o p a t i e n t s o f G r - 1 a n d Gr-2 f o r 4 months and that of Gr-3 for 1 year and 8 months. R e s u l t s : 1) I n b o t h Gr-1 and G r - 2 , serum T.G. LDL, VLDL, AST, ALT, G-GTF, a s w e l l a s l l p i d p e r o x i d e were h i g h e r t h a n normal level. Simvastatin treatment could lower those c h a n g e s b y 10-30%. 2) Hepatic CT v a l u e a t S 7 was r e c o v e r e d hy Simvastatin in Gr-1. (initial vales vs treated value: 5 5 . 0 ± 3 . 6 HU a n d 6 5 . 7 ~ 2 . 4 HU, p < 0 . 0 5 , r e s p e c t i v e l y ) 3) In Gr-3, Simvasta%in could improve liver pathology ( s w e l l i n g a n d n e c r o s i s o f h e p a t o e y t e s , PMN a n d l y m p h o c y t e s inf$1tration) and hyperlipidemia. C o n c l u s i o n : These d a t a indicate continuously drinking inducing alcoholic fatty liver and hepatitis with hyperlipidemia and lipidperoxidation, may lead to liver cell injury. S i m v a s t a t i n improved t h o s e c h a n g e s i n a l c o h o l i c f a t t y l i v e r a n d h e p a t i t i s w i t h a n d w i t h o u t HCV i n f e c t i o n .
DETECTION OF HEPATITIS C VHiUS USING AN AUTOMATED PCR INSTRUMENT SYSTEM. C. Young. S. Barmat, T. Haley, S. Lombardi. and L. Wolfe. Roche Molecular Systems, Inc., Branchburg, New Jersey, 08876-1760 Three hundred thirty eight serum or plasma specimens were tested for the presence of Hepatitis C Vires (HCV) using a prototype HCV Polymerase Chain Reaction (PCR) assay on an automated instrument system (COBAS® AMPLICORTM). The COBAS AMPLICOR Instrument System contains two independently controlled thermal eyelet blocks, a robotic arm for reagent handling, and a flexible analyzer for detection, thereby, integrating amplification, detection, and data/reagent management into an automated format that provides "walk away" capability to the user. The prototype HCV PCR assay was a coamplification of HCV and an internal control (IC) designed to monitor the amplification capability of each processed specimen. Sensitivity testing using RNA transcript demonstrated that the system was capable of detecting 5 input copies ofHCV RNA 100% of the time and inchisivity studies have shown that the assay detects all common HCV genotypes. The clinical specimens were evaluated by the automated prototype HCV assay in a blind study and the results were compared to the AMPLICORT M HCV Test in the mierowell plate format. When available, EL& and RIBA testing were used to resolve discrepant results bctween the two tests. Ofthe 222 AMPLICOR HCV negative samples tested, there was one false positive result using the prototype HCV test, for a specificity of 99:5% for this sample population: All 116 HCV positive samples were positive using the prototype HCV test, for a sensitivity of 100% enmpared to the AMPLICOR HCV Test. Overall, there was a 98.8% concordance between the AMPLICOR HCV Test and the prototype automated assay during initial testing and 100% concordance upon repeat testing. Ofthe 338 specimens tested, 3 had low IC values requiring repeat extraction and testing. Upon repeat, all 3 IC values recovered to normal levels. None of the 338 clinical specimens were determined to be PCR inhibited based on IC results. Overall, the prototype PCR assay on the COBAS AMPLICOR Instrument System performed equivalently to the AMPLICOR HCV Test in the microwell plate format for the deteetien of HCV in human serum or plasma.
It has been proven that interferon alpha 2a is effective in the treatment of chronic hepatitis C(HC), however the appropriate dosage and sehednle in Chinese patients remaines to be determined. The object of this study was to compare the efficacy of a dose of 3 MU of Roferon three times a week for 6 months with a starting dose of 6 MU for 3 months and a subsequent reduction to 3 MU three times a week for further 3 months. Sixty-eight patients with serological and histological proof of chronic hepatitis C with elevated serum ALl" levels were randomized into tWOgroups. 63 patients completed the full course of treatment. 5 patients were withdrawn from the study, 4 due to personal reasons and one due to aggravation of disease during treatment. 34 patients received 3 MU Referen t.i.w, for 6 months (group A), and 29 patients received 6 MU Roferon t.i.w. 3 months followed by 3 MU t.i.w, for another 3 months (group B). The sex, age, serum bilirubin, ALT and AST levels were matched in both groups. At the end of the 6th month the complete response and: partial response rates in group A were 67.6% and 8.8% respectively, the clearance of serum HCV-RNA was 71.4%. In group B, the complete and partial response rates were 62% and 20.7% and the elearanee of HCV-RNA was 72%. The efficacy between two dosage groups showed no statistical difference. The common adverse reactions (ADRS) were also similar in both groups. Most ADRS were tolerable only one patient suffered severe leucopenia requiringdose reduction. Another one patient suffered from marked elevation of serum bilirobin and ALT and aggravation of clinical symptoms, who was withdrawn from treatment. The response rates of Roferon treatment were correlated with genotype of HCV, severity of histopatho!ogy and titer of antibiodies to interferon.
1623
HEPATOLOGY O c t o b e r 1 9 9 5
1624
QUANTITATION OF ANTI-CORE HCV ANTIBODIES IS USEFUL FOR MONITORING IFN-THERAPY IN C H R O N I C HEPATITIS C. R Zachoval, R Hoffmann, G Michel, W Swoboda, H Dieoolder. MC Juno. G Frrsner and G R Pape. Medizinische Klinik II, Klinikum G-roBhadern and M a x von Pettenkofer-Institut, University o f Munich; Abbott Laboratories, Wiesbaden; Germany. Interferon.alpha (IFN) is an effective treatment in 20-30% o f patients with chronic hepatitis C (cHC). Evaluation o f effectiveness is based on serial alanine aminotransferase (ALT) measurements and determination o f H C V R N A in the serum. Serological markers o f efficacy are badly needed because they better reflect the patient's immune response to H C V and therapy. We therefore evaluated the usefulness o f quantitating antibodies to H C V core o f the I g G and separately o f the I g M type (automated immunoassays on the IMx analyser, Abbott Laboratories) during the course o f IFN therapy and up to at least 6 months after t h e end o f treatment. Thirty-six patients were included in the study: 10/36 were sustained responders (SR) to a course o f 3-5 Mio IFN s.c., thrice weekly for 6-12 months whereas 14 patients were classified as transient responders (TR) and 12 individuals as nonresponders (NR). The titer o f anti-HCV core IgM and I g G declined significantly (p<0.05, MarmWhitney U-test) at the end o f treatment in TR and SR compared to NR. Clinical relapse in TR was associated with an early increase o f both types o f specific antibodies whereas they remained at a constant low level in SR. In our study population we were not able to demonstrate a correlation between the total amount o f antibodies against H C V core before treatment and sustained response or the viral load in the serum (Amplicor Monitor, Hoffmarm-La Roche, Switzerland). Our data demonstrate that sequential quantitative assessment o f antiH C V core I g G and IgM is a valuable tool in monitoring responsiveness to IFN therapy in eriC.