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Relationship between HCV genotypes and routes of contamination in patients with chronic hepatitis C
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QUANTITATION OF HCV GENOME MOLECULES PATIENTS WITH LKM-I POSITIVE CHRONIC HEPATITIS
IN
F. Giostrn~ *A. Manzin I R. Francesconi, M. Lenzi, *L. Solforosi, *M. Clementl, F.B. Binnchl. Cattedra di Medicine Interoa I, Unlversltb dl Bologna. * Istituto di Mlcrobinlogin, Unlversitb dl Ancona. Italy. The majority of adult LKM-I positive patients wlth CH is also positive for anti?HCV antibodies end seror, HCV RNA. A pathogenetlc role of the LKM-I antibody. Is at present under reevaluation since It has been recently shown that Its target antigen (P450IID6) may be expressed on the hapatocellular plasmamembrana. As HCV infection is concerned, it has been shown that the prevalent HCV genotype is G1 according to Simmonds. Information is not avaible on the quantitation of HCV vi~mia. We have analized 21 patients, 10 positive for LKM-1 antibody and 11 negative. All were positive for anti-HCV and plasma HCV RNA by PCR. LKM-1 positive patients (SF/2M) had a median age of 57 years (range 22-83), median ALT 4.4X upper normal limit (range 1.4-1.7) and a liver histology with a mild/moderate activity in 9 subjects. Two LKM-1 positive patients had been treated with steroids. The 11 LKM-1 negative patients (2F/gM) had a median age of 40 (range 40-59), ALT values 2.7X upper normal limit (range 111.4) and all had liver histology with mild/moderate activity. Competitive RT.PCR was performed in two steps, cDNA was synthetized by using an antisense primer specific for genomic HCV RNA strand in the presence of HCV genomic sequences purified from plasma samples and a known amount of RNA competitor. The PCR products were scanned using a video densitometer. LKM-Uanti HCV positive patients showed a lower median value of HCV genome molecules (175788/ml, range 9780-1309347) than LKM-I negative/anti HCV positive ones (2885465/ml, range 331541-51675059) (p<0.0002). LKM-1/anti HCV positive patients show significantly lower levels of HCV viremin than the control group. The two LKM-1 positive with the highest values were those previously treated with steroids. The presence of low viremla associated with higher values of transaminuses suggests a concomitant autoimmune mechanism.
DETECTION OF HCV-RNA: A COMPARISON OF HCV-RNA ASSAYS. T.Cuypers z , M. Damen z , H,Reesink 2 . H.Zaaijer z . N.Lelie z. Central Lab. of the Blood Transfusion Service z and Red Cross Blood Bank 2, Amsterdam, The Netherlands. We compared the performance of the Roche Amplicor HGV-RNA PCR assay (Ampl.) for HCV-RNA detection in plasma with a well-validated "in house" cDNA-PCR test and the bDNA assay (Chiron Corp.). Three panels of samples were analyzed; 80 anti-HCV (RIBA) pos. samples and 42 anti-HCV neg. samples; two dilution series from the ist Eurohep reference pane'l (ZaalJer et al. Lancet 1993; 341:722); and two dilution series of genoUyped plasma's (type I and 3). Results; sensitivity and specificity. assay Anti-HCV+ (n-80) anti-HCVAmpl. 71/80 (88.8%) cDNA-PCR 72/80 (90.0%) 0 bDNA 54/80 (67.5%) 0 Results; reciprocal end-point titers series, assay Euroh-i Euroh-2 genot.-I Ampl. 5500 i000 4000 PCR I0,000 i0,000 64,000
(n-42). 0
in
dilution genot.-3 i000 16,000
We conclude, that the Amplicor HCV-RNA test has good specificity, but is approximately i log less sensitive as compared to our "in house" eDNA-PCR test. The HCVAmpllcor system is a simple and easy to perform HCVRNA test, that can be carried out in less than 6 h, whereas the "in house" assay needs three days before the results become available.
[ P1 Cl/64 I HCV gen0type II is associated to more severe liver disease and 10wer response to IFN treatment in Spanish patients C. Perez, F. Miralles~ O. Torras, R. Semina9or J. 8oadasr J. Pamplona~ JF. Sancho Poch¢, J. [nriquez. Onidad de Hepatologia. 5ervicio de Patologia Oigestiva. ¢Servicio de Anatomia Patol6gica. Hospital oe ,a Santa Creu i Sant Pau. Barcelona, Spain. Several genotypes of HCV have been recently identified, with a variable geographic distribution. Oifferent genotypes seem to be associated with the evolution of hepatitis C virus and with the response to IFN therapy. The aim of this study was to investigate the prevalence of HCV genotypes among anti-HCV and RNA-HCV positives patientswith different histologic damage. Patients: Group I: 15 blood donors with normal ALT levels. Group 2:24 ~with chronic persistent hepatitis (CPH). Group 3:44 patients with CAH. Group 4:25 patients with cirrhosis. Group 5:14 patients with HCC. Eigteen six of I08 patients of groups 2, 3, 4,-were treated with alpha-IFN, 48 of these being long responders at the end of therapy. None ~as addict to parenteral drugs. Methods: HCV RNA was detected using nested PCR and primers located in te-l~'e-5~noncoding region. Genotyping:nested PCR with specific primers localed in the capsid region (Okamotoet al,J Gen Virol 1992;73:673-9). Results: Genotype II was more prevalent in patients with HCC: Group I ~r37i'5")'~" group 2 (9/24); group 3 (23/44); group 4 (20/25); group S (II/14), (p
R E L A T I O N S H I P BETWEEN HCV GENOTYPES A N D ROUTES OF C O N T A M I N A T I O N IN P A T I E N T S W I T H C H R O N I C HEPATITIS C IM. Pawlotsky. L. Tsakiris.F, Roudot-Thoraval. C. Pellet.L. Stuyver, I. Dural, D. Dh~meau~c C H U Henri Mondor-Universit6 Paris XII, Cr~teil, France and Innogenetics, Ghent, Belgium. The aim of this work was to study the relationship between the H C V genotypes and the routes of viral contamination in a French series of unselected patients with chronic hepatitisC. Methods :101 patients with chronic hepatitisC (66 men, 35 women, mean age 47 yr) were studied.H C V genotype was determined by the Inno-LiPA H C V kit.The causes of H C V contamination were sought. Results : The distribution of H C V genotypes in the 101 • T~. [ ] Typee 2=,,41S patients was : la=20%, 1b=43%, per .cent U r~,lb 2a=10%, 3a=20%, 4 or 5a=7%. T. . . . Sbcty-four patients (63%) had an 1 i d e n t i f i e d cause of H C V transmission : b l o o d transfusion (BT) in 43 (43%) and intravenous drag use (IVDU) in 21 (21%). The remaining 37% had a =community-acquired" (CA) chronic hepatitis C. I V D U were IVDU BT CA significantly(p<0.0001)younger than B T and C A (33 and 51 yr, respectively).As shown in the figure. the distribution of H C V genotypes was similar in BT and CA, and significantlydifferentbetween I V D U and B T / C A (p<0.0001). Conclusions :In French patients with chronic hepatitisC, IVD U are younger and mainly infected with H C V genotype 3a. Patients with BT and C A have similar features,i.e,an older age and H C V Ib as main genotype. Our resultssuggest that two independent epidemics of hepatitis C are simuRaneously ongoing in France, with few crosscontaminations between the two epidemiological groups.
$38
QUANTITATION OF HCV GENOME MOLECULES PATIENTS WITH LKM-I POSITIVE CHRONIC HEPATITIS
IN
F. Giostrn~ *A. Manzin I R. Francesconi, M. Lenzi, *L. Solforosi, *M. Clementl, F.B. Binnchl. Cattedra di Medicine Interoa I, Unlversltb dl Bologna. * Istituto di Mlcrobinlogin, Unlversitb dl Ancona. Italy. The majority of adult LKM-I positive patients wlth CH is also positive for anti?HCV antibodies end seror, HCV RNA. A pathogenetlc role of the LKM-I antibody. Is at present under reevaluation since It has been recently shown that Its target antigen (P450IID6) may be expressed on the hapatocellular plasmamembrana. As HCV infection is concerned, it has been shown that the prevalent HCV genotype is G1 according to Simmonds. Information is not avaible on the quantitation of HCV vi~mia. We have analized 21 patients, 10 positive for LKM-1 antibody and 11 negative. All were positive for anti-HCV and plasma HCV RNA by PCR. LKM-1 positive patients (SF/2M) had a median age of 57 years (range 22-83), median ALT 4.4X upper normal limit (range 1.4-1.7) and a liver histology with a mild/moderate activity in 9 subjects. Two LKM-1 positive patients had been treated with steroids. The 11 LKM-1 negative patients (2F/gM) had a median age of 40 (range 40-59), ALT values 2.7X upper normal limit (range 111.4) and all had liver histology with mild/moderate activity. Competitive RT.PCR was performed in two steps, cDNA was synthetized by using an antisense primer specific for genomic HCV RNA strand in the presence of HCV genomic sequences purified from plasma samples and a known amount of RNA competitor. The PCR products were scanned using a video densitometer. LKM-Uanti HCV positive patients showed a lower median value of HCV genome molecules (175788/ml, range 9780-1309347) than LKM-I negative/anti HCV positive ones (2885465/ml, range 331541-51675059) (p<0.0002). LKM-1/anti HCV positive patients show significantly lower levels of HCV viremin than the control group. The two LKM-1 positive with the highest values were those previously treated with steroids. The presence of low viremla associated with higher values of transaminuses suggests a concomitant autoimmune mechanism.
DETECTION OF HCV-RNA: A COMPARISON OF HCV-RNA ASSAYS. T.Cuypers z , M. Damen z , H,Reesink 2 . H.Zaaijer z . N.Lelie z. Central Lab. of the Blood Transfusion Service z and Red Cross Blood Bank 2, Amsterdam, The Netherlands. We compared the performance of the Roche Amplicor HGV-RNA PCR assay (Ampl.) for HCV-RNA detection in plasma with a well-validated "in house" cDNA-PCR test and the bDNA assay (Chiron Corp.). Three panels of samples were analyzed; 80 anti-HCV (RIBA) pos. samples and 42 anti-HCV neg. samples; two dilution series from the ist Eurohep reference pane'l (ZaalJer et al. Lancet 1993; 341:722); and two dilution series of genoUyped plasma's (type I and 3). Results; sensitivity and specificity. assay Anti-HCV+ (n-80) anti-HCVAmpl. 71/80 (88.8%) cDNA-PCR 72/80 (90.0%) 0 bDNA 54/80 (67.5%) 0 Results; reciprocal end-point titers series, assay Euroh-i Euroh-2 genot.-I Ampl. 5500 i000 4000 PCR I0,000 i0,000 64,000
(n-42). 0
in
dilution genot.-3 i000 16,000
We conclude, that the Amplicor HCV-RNA test has good specificity, but is approximately i log less sensitive as compared to our "in house" eDNA-PCR test. The HCVAmpllcor system is a simple and easy to perform HCVRNA test, that can be carried out in less than 6 h, whereas the "in house" assay needs three days before the results become available.
[ P1 Cl/64 I HCV gen0type II is associated to more severe liver disease and 10wer response to IFN treatment in Spanish patients C. Perez, F. Miralles~ O. Torras, R. Semina9or J. 8oadasr J. Pamplona~ JF. Sancho Poch¢, J. [nriquez. Onidad de Hepatologia. 5ervicio de Patologia Oigestiva. ¢Servicio de Anatomia Patol6gica. Hospital oe ,a Santa Creu i Sant Pau. Barcelona, Spain. Several genotypes of HCV have been recently identified, with a variable geographic distribution. Oifferent genotypes seem to be associated with the evolution of hepatitis C virus and with the response to IFN therapy. The aim of this study was to investigate the prevalence of HCV genotypes among anti-HCV and RNA-HCV positives patientswith different histologic damage. Patients: Group I: 15 blood donors with normal ALT levels. Group 2:24 ~with chronic persistent hepatitis (CPH). Group 3:44 patients with CAH. Group 4:25 patients with cirrhosis. Group 5:14 patients with HCC. Eigteen six of I08 patients of groups 2, 3, 4,-were treated with alpha-IFN, 48 of these being long responders at the end of therapy. None ~as addict to parenteral drugs. Methods: HCV RNA was detected using nested PCR and primers located in te-l~'e-5~noncoding region. Genotyping:nested PCR with specific primers localed in the capsid region (Okamotoet al,J Gen Virol 1992;73:673-9). Results: Genotype II was more prevalent in patients with HCC: Group I ~r37i'5")'~" group 2 (9/24); group 3 (23/44); group 4 (20/25); group S (II/14), (p
R E L A T I O N S H I P BETWEEN HCV GENOTYPES A N D ROUTES OF C O N T A M I N A T I O N IN P A T I E N T S W I T H C H R O N I C HEPATITIS C IM. Pawlotsky. L. Tsakiris.F, Roudot-Thoraval. C. Pellet.L. Stuyver, I. Dural, D. Dh~meau~c C H U Henri Mondor-Universit6 Paris XII, Cr~teil, France and Innogenetics, Ghent, Belgium. The aim of this work was to study the relationship between the H C V genotypes and the routes of viral contamination in a French series of unselected patients with chronic hepatitisC. Methods :101 patients with chronic hepatitisC (66 men, 35 women, mean age 47 yr) were studied.H C V genotype was determined by the Inno-LiPA H C V kit.The causes of H C V contamination were sought. Results : The distribution of H C V genotypes in the 101 • T~. [ ] Typee 2=,,41S patients was : la=20%, 1b=43%, per .cent U r~,lb 2a=10%, 3a=20%, 4 or 5a=7%. T. . . . Sbcty-four patients (63%) had an 1 i d e n t i f i e d cause of H C V transmission : b l o o d transfusion (BT) in 43 (43%) and intravenous drag use (IVDU) in 21 (21%). The remaining 37% had a =community-acquired" (CA) chronic hepatitis C. I V D U were IVDU BT CA significantly(p<0.0001)younger than B T and C A (33 and 51 yr, respectively).As shown in the figure. the distribution of H C V genotypes was similar in BT and CA, and significantlydifferentbetween I V D U and B T / C A (p<0.0001). Conclusions :In French patients with chronic hepatitisC, IVD U are younger and mainly infected with H C V genotype 3a. Patients with BT and C A have similar features,i.e,an older age and H C V Ib as main genotype. Our resultssuggest that two independent epidemics of hepatitis C are simuRaneously ongoing in France, with few crosscontaminations between the two epidemiological groups.