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Difficulties in serological genotyping of HCV infection in patientswith porphyria cutanea tarda
182
Puppers read by title
DECORIN EXPRESSION AND DISTRIBUTION IN CHRONIC HEPATITIS C K. J&mav. G. Kar&sonv. M. Gallai*. I. Naav. Zs. Schti 1st Dept. Internal Medicine, Szent-Gybrgyi Medical University, Steged, *Ist Institute of Pathology yd Experimental Cancer Research, Semmelweis Me&al University Budapest, Hungary.
HEPATITIS DELTA GENOTYPES IN CHRONIC IN THE NORTHEAST OF SPAIN M.BUTI,
1
tin::M
AND
SENSITIVITY
SAMPLES
:
STUDY
U24 et service d’Hkpatologie,
Hdpital Beaujon,
Clichy,
The biNA signal amplification assay albws an accurate quantification of serum HCV RNA, with a lower sensitivii than PCR. Nevertheless, some studies have reported a lack of specificity of bDNA assay within the range of 0.2 to 0.6 mEq genomes/ml. The aim of our study was to evaluate : l)-the specificity 2)- the sensitivity of this assay. Patients and Methods : For the specificity study. 122 anti-HCV negative, 42 Hbs Ag positive, 42 anti-HCV positive/PCR negative and 70 with a low titer (0,2-0,6 mEq /ml) patients were studied For the sensitivity study 115 patients with a chronic hepatitis C included in a controlled trial of alpha interferon were studied. Serum HCV RNA quantification was performed with the bDNA signal amplification assay (HCV Quantiplex, Chiron, France), and the HCV Monitor assay (Roche, France) and Amplicor (Roche, France) for the qualitative PCR was used. Results: SDecificitv 0.2 to 0.6 Anti-HCV HBS Aa + Anti-HCV + PCR mEq /ml 90% Specificity 96% 100% 100% Among the 115 patients with chronic hepatitis C, 95 (63%) and 112 (96%) had detectable serum HCV RNA with Quantiplex and Monitor assays respectivelv. The mean serum HCV RNA levels were 3.97’578 mEq/ml-and 0.29f0.69 RNA copies/ml with the two assays respectively. The results obtained with the two assays were not correlated (r=O.401). Conclusions. The overall specificity of the bDNA signal amplification assay was 96%. In patients with chronic hepatitis C, as compared with qualitative PCR, the sensitivity of the bDNA assay was 63%. There was no relationship between results obtained by the Quantiplex and the Monitor assays. The respective sensitivity and the absence of correlation between the two assays interprete clinical studies.
should
be taken
into account
F.RODRIGUEZ,
to
J.CASTELLS,
Barcelona,
Spain.
Hepatitis delta virus IHDV) genotypes have been associated with different disease patterns and geographic distribution. To determine the prevalence of HDV genotypes in Spain and the correlation with transmission routes and clinical disease the nucleotide divergence of the consensus sequence of HDV-RNA were studied from 33 patients with chronic delta hepatitis (24 iv. drug users and 9 without risk factors) and 4 patients with acute self-limited delta hepatitis. Serum HDV-RNA was amplified by PCR and a fragment of 350 nucleotides was directly sequenced. Genetic analysis of the consensus sequence obtained showed a high degree of conservation between nucleotides sequence (mean 93%). The comparison of these sequences with those derived from different geographic areas and pertaining to genotypes I, II and Ill showed a mean sequence identity of 92% with genotype I, 73% with genotype II and 61% with genotype Ill. No HDV subgenotypes were discernable. There was no association between the type of infection and the distribution of sequences on the phylogenetic tree generated. In conclusion, these results indicate that HDV isolates in our area are exclusively genotype I, independently of the transmission route and the type of infection and suggest an homogeneous origin of HDV infection in our area.
1 CO5/008
MEASURINGSERUM HCV RNA IN CLINICAL A SPECIFICITY
M.COTRINA,
J.QUER, R. ESTEBAN, J. GUARDIA. Liver Unit. Hospital Universitario Valle Hebr6n.
Backwound/Aims: Decorin is a small leucine-rich extracellular matrix proteoglycan with significance in regulation of fibrogenesis. In this study were assessed the distribution of its expression in chronic hepatitis C (CH-C) before and after interferon-cc (m-x) treatment. Patients and methods: 20 patients (13 men, 7 women) with CH-C received FN-oc therapy (3x3 MU TIW) over 6 months (n=lO) and over 12 months (n=lO). Liver biopsy samples were taken before and after IFN-oc treatment using a monoclonal antibody with tyramin amplification and avidin-biotin-peroxidase immunohistochemical method for detection of the decorin. Statistical analysis was performed with Wilcoxon signed rank test and with Spearman rank order test. Results: Decorin deposition increased parallel with the inflammatory activity (grade) of CH-C. In severe active hepatitis it was detected not only in the centrilobular and in the periportal regions - as in normal cases - but in the perisinusoidal areas and as bridging reactions between the portal tracts. IFN-a treatment reduced significantly the amount of the deposited decorin as well as the necroinflammatory activity (grade) of CHC in the 12 months cohort. Conclusions: The long-term IFN-oc therapy decreased the decorin deposition in CH-C.
1 COW06
R. JARDI,
DELTA INFECTION
1
DIFFICULTIES IN SEROLOGICAL GENOTYPING OF HCV INFECTION IN PATIENTS WITH PORPHYRIA CUTANEA TAFtDA Clinic of Gastroenterology, Bulgaria.
St. Ivan Rilski’ University
Hospital,
Sofia,
In many countries the prevalence of HCV infection among the patients with porphyria cutanea tarda (PCT) is extremely high. The present investigation aimed to compare the distribution of HCV genotypes in PCT patients with chronic HCV infection and HCV individuals without PCT. Patients: In 36 of 70 PCT patients HCV-antibodies were established using ELISA III generation with confirmation by Western blot assay. In another 45 individuals without PCT a chronic HCV infection was proven by establishing HCV-antibodies and the presence of HCV RNA (RTPCR). In both groups the genotypes were investigated by Murex HCV Serotyping 1-6 Assay. It is based on the positive response of antisera to antigenic sites of viral NS4 protein. Re,sul& Among the 36 anti-HCV positive PCT patients antibodies against NS4 region were not detected in 14 (38,9%). This precluded genotyping in these persons. The other 22 PCT patients were found positive for NS4 antibodies. Three of them were untypable. All the remaining 19 patients belonged to genotype 1. In addition, 3 of them had also genotype 2 and 1 individual had genotype 3. Unlike PCT group, NS4 antibodies were found in all 45 persons without PCT. Genotype 1 alone was established in 35 of them and genotype 2 alone in 2. r&infection was found in 4 patients - genotypes 1+2 in 1, genotypes 2+3 in 1 and genotypes 1+2+3 in 2. Four patients remained untypable. Conclusions: Genotype 1 was the most frequent one among the studied series of HCV patients including the individuals with PCT. However, the serological genotyping does not seem suitable in PCT patients due to the lack of antibodies against NS4 region in considerable number of them. The reasons and significance of this obstacle remain obscure and deserve further investigation.
Puppers read by title
DECORIN EXPRESSION AND DISTRIBUTION IN CHRONIC HEPATITIS C K. J&mav. G. Kar&sonv. M. Gallai*. I. Naav. Zs. Schti 1st Dept. Internal Medicine, Szent-Gybrgyi Medical University, Steged, *Ist Institute of Pathology yd Experimental Cancer Research, Semmelweis Me&al University Budapest, Hungary.
HEPATITIS DELTA GENOTYPES IN CHRONIC IN THE NORTHEAST OF SPAIN M.BUTI,
1
tin::M
AND
SENSITIVITY
SAMPLES
:
STUDY
U24 et service d’Hkpatologie,
Hdpital Beaujon,
Clichy,
The biNA signal amplification assay albws an accurate quantification of serum HCV RNA, with a lower sensitivii than PCR. Nevertheless, some studies have reported a lack of specificity of bDNA assay within the range of 0.2 to 0.6 mEq genomes/ml. The aim of our study was to evaluate : l)-the specificity 2)- the sensitivity of this assay. Patients and Methods : For the specificity study. 122 anti-HCV negative, 42 Hbs Ag positive, 42 anti-HCV positive/PCR negative and 70 with a low titer (0,2-0,6 mEq /ml) patients were studied For the sensitivity study 115 patients with a chronic hepatitis C included in a controlled trial of alpha interferon were studied. Serum HCV RNA quantification was performed with the bDNA signal amplification assay (HCV Quantiplex, Chiron, France), and the HCV Monitor assay (Roche, France) and Amplicor (Roche, France) for the qualitative PCR was used. Results: SDecificitv 0.2 to 0.6 Anti-HCV HBS Aa + Anti-HCV + PCR mEq /ml 90% Specificity 96% 100% 100% Among the 115 patients with chronic hepatitis C, 95 (63%) and 112 (96%) had detectable serum HCV RNA with Quantiplex and Monitor assays respectivelv. The mean serum HCV RNA levels were 3.97’578 mEq/ml-and 0.29f0.69 RNA copies/ml with the two assays respectively. The results obtained with the two assays were not correlated (r=O.401). Conclusions. The overall specificity of the bDNA signal amplification assay was 96%. In patients with chronic hepatitis C, as compared with qualitative PCR, the sensitivity of the bDNA assay was 63%. There was no relationship between results obtained by the Quantiplex and the Monitor assays. The respective sensitivity and the absence of correlation between the two assays interprete clinical studies.
should
be taken
into account
F.RODRIGUEZ,
to
J.CASTELLS,
Barcelona,
Spain.
Hepatitis delta virus IHDV) genotypes have been associated with different disease patterns and geographic distribution. To determine the prevalence of HDV genotypes in Spain and the correlation with transmission routes and clinical disease the nucleotide divergence of the consensus sequence of HDV-RNA were studied from 33 patients with chronic delta hepatitis (24 iv. drug users and 9 without risk factors) and 4 patients with acute self-limited delta hepatitis. Serum HDV-RNA was amplified by PCR and a fragment of 350 nucleotides was directly sequenced. Genetic analysis of the consensus sequence obtained showed a high degree of conservation between nucleotides sequence (mean 93%). The comparison of these sequences with those derived from different geographic areas and pertaining to genotypes I, II and Ill showed a mean sequence identity of 92% with genotype I, 73% with genotype II and 61% with genotype Ill. No HDV subgenotypes were discernable. There was no association between the type of infection and the distribution of sequences on the phylogenetic tree generated. In conclusion, these results indicate that HDV isolates in our area are exclusively genotype I, independently of the transmission route and the type of infection and suggest an homogeneous origin of HDV infection in our area.
1 CO5/008
MEASURINGSERUM HCV RNA IN CLINICAL A SPECIFICITY
M.COTRINA,
J.QUER, R. ESTEBAN, J. GUARDIA. Liver Unit. Hospital Universitario Valle Hebr6n.
Backwound/Aims: Decorin is a small leucine-rich extracellular matrix proteoglycan with significance in regulation of fibrogenesis. In this study were assessed the distribution of its expression in chronic hepatitis C (CH-C) before and after interferon-cc (m-x) treatment. Patients and methods: 20 patients (13 men, 7 women) with CH-C received FN-oc therapy (3x3 MU TIW) over 6 months (n=lO) and over 12 months (n=lO). Liver biopsy samples were taken before and after IFN-oc treatment using a monoclonal antibody with tyramin amplification and avidin-biotin-peroxidase immunohistochemical method for detection of the decorin. Statistical analysis was performed with Wilcoxon signed rank test and with Spearman rank order test. Results: Decorin deposition increased parallel with the inflammatory activity (grade) of CH-C. In severe active hepatitis it was detected not only in the centrilobular and in the periportal regions - as in normal cases - but in the perisinusoidal areas and as bridging reactions between the portal tracts. IFN-a treatment reduced significantly the amount of the deposited decorin as well as the necroinflammatory activity (grade) of CHC in the 12 months cohort. Conclusions: The long-term IFN-oc therapy decreased the decorin deposition in CH-C.
1 COW06
R. JARDI,
DELTA INFECTION
1
DIFFICULTIES IN SEROLOGICAL GENOTYPING OF HCV INFECTION IN PATIENTS WITH PORPHYRIA CUTANEA TAFtDA Clinic of Gastroenterology, Bulgaria.
St. Ivan Rilski’ University
Hospital,
Sofia,
In many countries the prevalence of HCV infection among the patients with porphyria cutanea tarda (PCT) is extremely high. The present investigation aimed to compare the distribution of HCV genotypes in PCT patients with chronic HCV infection and HCV individuals without PCT. Patients: In 36 of 70 PCT patients HCV-antibodies were established using ELISA III generation with confirmation by Western blot assay. In another 45 individuals without PCT a chronic HCV infection was proven by establishing HCV-antibodies and the presence of HCV RNA (RTPCR). In both groups the genotypes were investigated by Murex HCV Serotyping 1-6 Assay. It is based on the positive response of antisera to antigenic sites of viral NS4 protein. Re,sul& Among the 36 anti-HCV positive PCT patients antibodies against NS4 region were not detected in 14 (38,9%). This precluded genotyping in these persons. The other 22 PCT patients were found positive for NS4 antibodies. Three of them were untypable. All the remaining 19 patients belonged to genotype 1. In addition, 3 of them had also genotype 2 and 1 individual had genotype 3. Unlike PCT group, NS4 antibodies were found in all 45 persons without PCT. Genotype 1 alone was established in 35 of them and genotype 2 alone in 2. r&infection was found in 4 patients - genotypes 1+2 in 1, genotypes 2+3 in 1 and genotypes 1+2+3 in 2. Four patients remained untypable. Conclusions: Genotype 1 was the most frequent one among the studied series of HCV patients including the individuals with PCT. However, the serological genotyping does not seem suitable in PCT patients due to the lack of antibodies against NS4 region in considerable number of them. The reasons and significance of this obstacle remain obscure and deserve further investigation.