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Epidemiology of hepatitis B and hepatitis C virus infection among HIV counselling and testing clients in Jos, north central Nigeria
Abstracts / Journal of Clinical Virology 69 (2015) 223–246
P0028
P0029
HBsAg decline after interrupting nucleos(t)ide analogue therapy in HVeAg-negative chronic hepatitis B
Are medical practitioners ready enough to deal with HCV in Karachi, Pakistan?
C. Höner zu Siederdissen 1 , F. Rinker 1,2 , C.S. Falk 3 , N. Filmann 4 , B. Maasoumy 1 , K. Deterding 1 , K. Port 1 , C. Mix 1 , E. Herrmann 4 , H. Wedemeyer 1,2 , M.P. Manns 1,2 , A.R.M. Kraft 1,2 , M. Cornberg 1,2,∗ 1
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany 2 German Center for Infection Research, Hannover, Germany 3 Institute of Transplant Immunology, IFB-Tx, Hannover Medical School, Hannover, Germany 4 Institute of Biostatistics and Mathematical Modeling, Faculty of Medicine, Goethe-University, Frankfurt am Main, Germany Background: HBsAg loss is rarely seen during treatment with nucleos(t)ide analogues (NA). Therefore, treatment is usually not finite and rules on when to stop are not clearly defined. Methods: In a prospective pilot trial, NA treatment was stopped in patients with HBeAg-negative chronic hepatitis B. Inclusion criteria were ongoing antiviral treatment, suppressed HBV DNA <20 IU/mL for at least 3 years, HBsAg <3500 IU/mL, and a verified indication for antiviral treatment at the time point of NA initiation, according to international guidelines. Patients were assessed for virological relapse, defined by HBV DNA >2000 IU/mL. Quantitative HBsAg levels 1 year before and 1 year after treatment cessation were compared to assess differences. Serum IP-10 (CXCL10) levels were analysed using multiplex technology. Findings: Data were available for 13 patients with 1 year followup. Virological relapse occurred in 12/13 patients (92%) within the first 6 months, primarily within the first 8 weeks. Interestingly, a significant increase in IP-10 levels could be observed 8 weeks after stopping NA therapy (p = 0.028). In 11/12 patients, treatment was re-initiated and HBV-DNA was successfully suppressed at week 48 follow-up. After treatment cessation, we detected a significant decline in median HBsAg levels from stopping NA therapy to 48 week follow-up (p = 0.0012), whereas there was no change during NA therapy 1 year before treatment cessation. ALT flare >5 ULN occurred in 2 patients and was associated with a >1 log HBsAg reduction. Interpretation: Treatment cessation in non-cirrhotic patients with HVeAg-negative chronic hepatitis B leads to virological relapse in >90% until week 24 but is associated with a significant HBsAg reduction after 1 year. Induction of serum IP-10 may help to explain the effect on HBsAg decline. Thus, interrupting NA treatment and inducing immune responses should be further investigated as an option to facilitate HBsAg loss. http://dx.doi.org/10.1016/j.jcv.2015.06.035
233
S. Hussain 1,∗ , K. Malik 1 , T. Niaz 1 , S. Zafar 1 , A. Kazi 1 , M. Munir 1 , O. Yacob 1 , R. Shehzad 1 , A. Alam 1 , S. Maan 1 , M. Ahmed 1 , N. Rizvi 2 1
The Aga Khan University, Karachi, Pakistan Department of Community Health Sciences, The Aga Khan University, Karachi, Pakistan 2
Background: Globally, 150–170 million people are chronic carriers of hepatitis C virus (HCV) and 350 000 deaths are attributed to it annually. HCV is the leading cause of cirrhosis and liver transplant. 10 million people are carriers of HCV in Pakistan, with prevalence between 2.2 and 14%. No vaccine is available. Healthcare workers constitute a high risk group as they are more likely to encounter patients with HCV during their clinical practice. HCV prevalence is 5.2% among health-care workers in Pakistan. The aim of this study is to determine knowledge and practices regarding HCV, its prevention, and treatment in medical practitioners in Karachi, Pakistan. Methods: 162 medical practitioners were surveyed in Karachi using a structured questionnaire. The respondents were interns, residents, and consultants, but excluded those working in hepatology, gastroenterology, and infectious diseases. Questions tested their knowledge of HCV in general, and practices according to WHO guidelines. The study was conducted during November and December, 2014. Findings: Among the respondents, 48.4% were male and 52.6% were female. 80.3% were under the age of 35. Cutoff was set at 70% for adequate knowledge, and at 50% for satisfactory practices. Knowledge regarding prevalence of HCV was inadequate, with 32.1% answering correctly. 66% thought that HCV should be tested for before invasive procedures. Almost 90% of doctors knew the correct routes of transmission of HCV. 72% knew that hepatitis is the most common cause of liver cirrhosis. 78.4% of doctors knew about extra-hepatic complications of HCV. 60.1% knew that pegylated interferon and ribavirin were treatment of choice. Less than 50% knew that aminotransferase and FIB-4 are the most appropriate tools for assessment of hepatic fibrosis. Interpretation: Knowledge about HCV was adequate in our respondents, as almost all questions had more than 70% correct responses. Adherence to WHO standards regarding practices was inadequate. Further medical education and reinforcement seminars need to be organised regularly for doctors practicing in Karachi. http://dx.doi.org/10.1016/j.jcv.2015.06.036 P0030 Epidemiology of hepatitis B and hepatitis C virus infection among HIV counselling and testing clients in Jos, north central Nigeria J.Y. Peter 1,∗ , O.O. Agbaji 1 , A.T. Olayinka 2 , F.T. Ogunsola 3 1
Jos University Teaching Hospital, Jos, Nigeria Ahmadu Bello University Teaching Hospital, Zaria, Nigeria 3 Lagos University Teaching Hospital, Lagos, Nigeria 2
Background: Viral hepatitis is common in Nigeria, where it is a major cause of both acute and chronic liver disease, as well as hepatocellular cancer. People at risk of acquisition of human
234
Abstracts / Journal of Clinical Virology 69 (2015) 223–246
immunodeficiency virus (HIV) infection are also at risk of acquisition of other blood-borne infections, including hepatitis B virus (HBV) and hepatitis C virus (HCV) infection. We set out to determine the epidemiology of HBV and HCV infection among HIV counselling and testing (HCT) clients at the Jos University Teaching Hospital (JUTH), Nigeria. Methods: This was a cross-sectional study conducted at the HCT unit of the AIDS Prevention Initiative in Nigeria (APIN), Jos University Teaching Hospital (JUTH), Jos, Nigeria, between November, 2012, and April, 2013. Patients were recruited consecutively at the HCT unit of APIN JUTH. Included patients were aged 18 years and above, who were antiretroviral (ARV) drug-naive and who accepted and signed the consent form. Clients who declined to sign the consent form were excluded. The study involved collecting demographic data, exposure to risk factors, and laboratory determination of HBV and HCV seroprevalence in the patients using ELISA and PCR assay methods. Chi-squared test was used to determine significance of association between categorical variables. Bivariate analysis was used to determine independent risk factors. Findings: 132 (56.9%) patients were women, and 100 (43.1%) were men. 36 (15.5%) tested positive for HBsAg by ELISA, and 31 (13.4%) of these patients were confirmed positive by DNA PCR. Nine (3.9%) tested positive by ELISA for HCV antibody, and seven (3.0%) were confirmed positive by RNA PCR. The coinfection rate of HIV and HBV was 5.2%. Infection was more common among those younger than 36 years, in the case of HBV, and those older than 36 years, in the case of HCV. Interpretation: We concluded the prevalence of HBV infection was high. The study was limited by the cross-sectional design. http://dx.doi.org/10.1016/j.jcv.2015.06.037 P0031 Deep sequencing shows microRNA expression in hepatitis C patients is associated with the end treatment response A.M. Butt 1,2,∗ , L. Castellano 2 , M. Idrees 1 , Y. Tong 3 1
University of the Punjab, Lahore, Pakistan Hammersmith Hospital, Imperial College London, UK 3 Beijing Institute of Microbiology and Epidemiology, Beijing, China 2
Background: Hepatitis C virus is a major cause of chronic liver disease worldwide. microRNAs (miRNAs) are small, non-coding RNA molecules that regulate gene expression. The impact of different miRNAs on HCV disease progression has been reported, but the data from such studies are restricted to analysis of a limited set of miRNAs. Recently, we reported next-generation sequencing (NGS) based profiling of differentially expressed miRNAs (DE-miRNAs) between patients with chronic hepatitis C (CHC) and healthy controls. In this study, we report identification via deep sequencing of DE-miRNAs between different groups of CHC patients grouped according to response to interferon therapy. Methods: Eight pretreatment liver biopsy specimens from CHC patients (sustained virological responders [SVR; n = 4] and nonresponders [NR; n = 4]) were selected for Illumina sequencing. DE-miRNAs were identified following NGS data analysis and validated by real-time PCR in an independent cohort of CHC patients (n = 120). The diagnostic and prognostic potential of DE-miRNAs in the different groups was assessed by receiver operating characteristic (ROC) curves and regression analyses. DE-miRNAs were also profiled in matched serum samples of cohort patients and controls, as non-invasive biomarkers.
Findings: We identified 23, 14, and eight DE-miRNAs between SVR:NR SVR:relapse, and NR:relapse patients groups (−1.5 < fold change > 1.5; P < 0.05), respectively. Expression of several hepatic and serum DE-miRNAs, including miR-122, -21, -221, and -125 correlated with biochemical and histological parameters and endtreatment response (ETR). An integrated miRNA–mRNA interaction analysis of DE-miRNAs showed that these miRNAs targeted several genes associated with important pathways, including interferon-␣, immune response, and lipid metabolism in the SVR and NR groups. Interpretation: The pretreatment levels of hepatic and serum DE-miRNAs correlated significantly with the ETR, highlighting the promising potential of these miRNAs as diagnostic biomarkers. Moreover, these miRNAs can also be exploited as novel therapeutics based on their impact on host gene expression and metabolic pathways. http://dx.doi.org/10.1016/j.jcv.2015.06.038 P0032 Detection of NS5B C316N—A resistant mutation to sofosbuvir in treatment-naïve genotype 1b HCV-infected patients using ultra-deep sequencing Y. Wang ∗ , H. Rao, L. Wei Peking University Hepatology Institute, Peking, China Background: The C316N mutation has been reported as the variant to impact the activity of certain NS5B-palm-targeting nonnucleoside polymerase inhibitors. Baseline N316 polymorphism was associated with treatment failure of the newest NS5B polymerase inhibitor sofosbuvir. We aimed to investigate whether this substitution pre-existed in treatment-naïve genotype 1b HCVinfected patients, and whether treatment of pegylated interferon (PegIFN) plus ribavirin (RBV) would impact the mutation frequencies. Methods: NS5B polymorphism deep sequencing was performed with an average coverage of 2000 in 24 genotype 1b HCV-infected patients both at baseline and after receiving therapy of PegIFN plus RBV for 48 weeks. Findings: C316N polymorphism was detected in 19 (79.2%) of 24 patients at baseline, with mutation frequencies of 6.2–93%. The prevalence was much higher than 12% in Europe, as reported by previous studies. When the treatment outcome was considered, no difference was found between patients who achieved sustained virological response (SVR) and who didn’t (75% vs 81.3%, p = 1.00). Treatment of PegIFN plus RBV for 48 weeks did not change the frequency of this mutation. Interpretation: The NS5B substitution C316N associated with treatment failure was identified with a quite high prevalence in this study. The detection of this mutation should be considered for patients in Asia receiving treatment of sofosbuvir in the near future. http://dx.doi.org/10.1016/j.jcv.2015.06.039
P0028
P0029
HBsAg decline after interrupting nucleos(t)ide analogue therapy in HVeAg-negative chronic hepatitis B
Are medical practitioners ready enough to deal with HCV in Karachi, Pakistan?
C. Höner zu Siederdissen 1 , F. Rinker 1,2 , C.S. Falk 3 , N. Filmann 4 , B. Maasoumy 1 , K. Deterding 1 , K. Port 1 , C. Mix 1 , E. Herrmann 4 , H. Wedemeyer 1,2 , M.P. Manns 1,2 , A.R.M. Kraft 1,2 , M. Cornberg 1,2,∗ 1
Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany 2 German Center for Infection Research, Hannover, Germany 3 Institute of Transplant Immunology, IFB-Tx, Hannover Medical School, Hannover, Germany 4 Institute of Biostatistics and Mathematical Modeling, Faculty of Medicine, Goethe-University, Frankfurt am Main, Germany Background: HBsAg loss is rarely seen during treatment with nucleos(t)ide analogues (NA). Therefore, treatment is usually not finite and rules on when to stop are not clearly defined. Methods: In a prospective pilot trial, NA treatment was stopped in patients with HBeAg-negative chronic hepatitis B. Inclusion criteria were ongoing antiviral treatment, suppressed HBV DNA <20 IU/mL for at least 3 years, HBsAg <3500 IU/mL, and a verified indication for antiviral treatment at the time point of NA initiation, according to international guidelines. Patients were assessed for virological relapse, defined by HBV DNA >2000 IU/mL. Quantitative HBsAg levels 1 year before and 1 year after treatment cessation were compared to assess differences. Serum IP-10 (CXCL10) levels were analysed using multiplex technology. Findings: Data were available for 13 patients with 1 year followup. Virological relapse occurred in 12/13 patients (92%) within the first 6 months, primarily within the first 8 weeks. Interestingly, a significant increase in IP-10 levels could be observed 8 weeks after stopping NA therapy (p = 0.028). In 11/12 patients, treatment was re-initiated and HBV-DNA was successfully suppressed at week 48 follow-up. After treatment cessation, we detected a significant decline in median HBsAg levels from stopping NA therapy to 48 week follow-up (p = 0.0012), whereas there was no change during NA therapy 1 year before treatment cessation. ALT flare >5 ULN occurred in 2 patients and was associated with a >1 log HBsAg reduction. Interpretation: Treatment cessation in non-cirrhotic patients with HVeAg-negative chronic hepatitis B leads to virological relapse in >90% until week 24 but is associated with a significant HBsAg reduction after 1 year. Induction of serum IP-10 may help to explain the effect on HBsAg decline. Thus, interrupting NA treatment and inducing immune responses should be further investigated as an option to facilitate HBsAg loss. http://dx.doi.org/10.1016/j.jcv.2015.06.035
233
S. Hussain 1,∗ , K. Malik 1 , T. Niaz 1 , S. Zafar 1 , A. Kazi 1 , M. Munir 1 , O. Yacob 1 , R. Shehzad 1 , A. Alam 1 , S. Maan 1 , M. Ahmed 1 , N. Rizvi 2 1
The Aga Khan University, Karachi, Pakistan Department of Community Health Sciences, The Aga Khan University, Karachi, Pakistan 2
Background: Globally, 150–170 million people are chronic carriers of hepatitis C virus (HCV) and 350 000 deaths are attributed to it annually. HCV is the leading cause of cirrhosis and liver transplant. 10 million people are carriers of HCV in Pakistan, with prevalence between 2.2 and 14%. No vaccine is available. Healthcare workers constitute a high risk group as they are more likely to encounter patients with HCV during their clinical practice. HCV prevalence is 5.2% among health-care workers in Pakistan. The aim of this study is to determine knowledge and practices regarding HCV, its prevention, and treatment in medical practitioners in Karachi, Pakistan. Methods: 162 medical practitioners were surveyed in Karachi using a structured questionnaire. The respondents were interns, residents, and consultants, but excluded those working in hepatology, gastroenterology, and infectious diseases. Questions tested their knowledge of HCV in general, and practices according to WHO guidelines. The study was conducted during November and December, 2014. Findings: Among the respondents, 48.4% were male and 52.6% were female. 80.3% were under the age of 35. Cutoff was set at 70% for adequate knowledge, and at 50% for satisfactory practices. Knowledge regarding prevalence of HCV was inadequate, with 32.1% answering correctly. 66% thought that HCV should be tested for before invasive procedures. Almost 90% of doctors knew the correct routes of transmission of HCV. 72% knew that hepatitis is the most common cause of liver cirrhosis. 78.4% of doctors knew about extra-hepatic complications of HCV. 60.1% knew that pegylated interferon and ribavirin were treatment of choice. Less than 50% knew that aminotransferase and FIB-4 are the most appropriate tools for assessment of hepatic fibrosis. Interpretation: Knowledge about HCV was adequate in our respondents, as almost all questions had more than 70% correct responses. Adherence to WHO standards regarding practices was inadequate. Further medical education and reinforcement seminars need to be organised regularly for doctors practicing in Karachi. http://dx.doi.org/10.1016/j.jcv.2015.06.036 P0030 Epidemiology of hepatitis B and hepatitis C virus infection among HIV counselling and testing clients in Jos, north central Nigeria J.Y. Peter 1,∗ , O.O. Agbaji 1 , A.T. Olayinka 2 , F.T. Ogunsola 3 1
Jos University Teaching Hospital, Jos, Nigeria Ahmadu Bello University Teaching Hospital, Zaria, Nigeria 3 Lagos University Teaching Hospital, Lagos, Nigeria 2
Background: Viral hepatitis is common in Nigeria, where it is a major cause of both acute and chronic liver disease, as well as hepatocellular cancer. People at risk of acquisition of human
234
Abstracts / Journal of Clinical Virology 69 (2015) 223–246
immunodeficiency virus (HIV) infection are also at risk of acquisition of other blood-borne infections, including hepatitis B virus (HBV) and hepatitis C virus (HCV) infection. We set out to determine the epidemiology of HBV and HCV infection among HIV counselling and testing (HCT) clients at the Jos University Teaching Hospital (JUTH), Nigeria. Methods: This was a cross-sectional study conducted at the HCT unit of the AIDS Prevention Initiative in Nigeria (APIN), Jos University Teaching Hospital (JUTH), Jos, Nigeria, between November, 2012, and April, 2013. Patients were recruited consecutively at the HCT unit of APIN JUTH. Included patients were aged 18 years and above, who were antiretroviral (ARV) drug-naive and who accepted and signed the consent form. Clients who declined to sign the consent form were excluded. The study involved collecting demographic data, exposure to risk factors, and laboratory determination of HBV and HCV seroprevalence in the patients using ELISA and PCR assay methods. Chi-squared test was used to determine significance of association between categorical variables. Bivariate analysis was used to determine independent risk factors. Findings: 132 (56.9%) patients were women, and 100 (43.1%) were men. 36 (15.5%) tested positive for HBsAg by ELISA, and 31 (13.4%) of these patients were confirmed positive by DNA PCR. Nine (3.9%) tested positive by ELISA for HCV antibody, and seven (3.0%) were confirmed positive by RNA PCR. The coinfection rate of HIV and HBV was 5.2%. Infection was more common among those younger than 36 years, in the case of HBV, and those older than 36 years, in the case of HCV. Interpretation: We concluded the prevalence of HBV infection was high. The study was limited by the cross-sectional design. http://dx.doi.org/10.1016/j.jcv.2015.06.037 P0031 Deep sequencing shows microRNA expression in hepatitis C patients is associated with the end treatment response A.M. Butt 1,2,∗ , L. Castellano 2 , M. Idrees 1 , Y. Tong 3 1
University of the Punjab, Lahore, Pakistan Hammersmith Hospital, Imperial College London, UK 3 Beijing Institute of Microbiology and Epidemiology, Beijing, China 2
Background: Hepatitis C virus is a major cause of chronic liver disease worldwide. microRNAs (miRNAs) are small, non-coding RNA molecules that regulate gene expression. The impact of different miRNAs on HCV disease progression has been reported, but the data from such studies are restricted to analysis of a limited set of miRNAs. Recently, we reported next-generation sequencing (NGS) based profiling of differentially expressed miRNAs (DE-miRNAs) between patients with chronic hepatitis C (CHC) and healthy controls. In this study, we report identification via deep sequencing of DE-miRNAs between different groups of CHC patients grouped according to response to interferon therapy. Methods: Eight pretreatment liver biopsy specimens from CHC patients (sustained virological responders [SVR; n = 4] and nonresponders [NR; n = 4]) were selected for Illumina sequencing. DE-miRNAs were identified following NGS data analysis and validated by real-time PCR in an independent cohort of CHC patients (n = 120). The diagnostic and prognostic potential of DE-miRNAs in the different groups was assessed by receiver operating characteristic (ROC) curves and regression analyses. DE-miRNAs were also profiled in matched serum samples of cohort patients and controls, as non-invasive biomarkers.
Findings: We identified 23, 14, and eight DE-miRNAs between SVR:NR SVR:relapse, and NR:relapse patients groups (−1.5 < fold change > 1.5; P < 0.05), respectively. Expression of several hepatic and serum DE-miRNAs, including miR-122, -21, -221, and -125 correlated with biochemical and histological parameters and endtreatment response (ETR). An integrated miRNA–mRNA interaction analysis of DE-miRNAs showed that these miRNAs targeted several genes associated with important pathways, including interferon-␣, immune response, and lipid metabolism in the SVR and NR groups. Interpretation: The pretreatment levels of hepatic and serum DE-miRNAs correlated significantly with the ETR, highlighting the promising potential of these miRNAs as diagnostic biomarkers. Moreover, these miRNAs can also be exploited as novel therapeutics based on their impact on host gene expression and metabolic pathways. http://dx.doi.org/10.1016/j.jcv.2015.06.038 P0032 Detection of NS5B C316N—A resistant mutation to sofosbuvir in treatment-naïve genotype 1b HCV-infected patients using ultra-deep sequencing Y. Wang ∗ , H. Rao, L. Wei Peking University Hepatology Institute, Peking, China Background: The C316N mutation has been reported as the variant to impact the activity of certain NS5B-palm-targeting nonnucleoside polymerase inhibitors. Baseline N316 polymorphism was associated with treatment failure of the newest NS5B polymerase inhibitor sofosbuvir. We aimed to investigate whether this substitution pre-existed in treatment-naïve genotype 1b HCVinfected patients, and whether treatment of pegylated interferon (PegIFN) plus ribavirin (RBV) would impact the mutation frequencies. Methods: NS5B polymorphism deep sequencing was performed with an average coverage of 2000 in 24 genotype 1b HCV-infected patients both at baseline and after receiving therapy of PegIFN plus RBV for 48 weeks. Findings: C316N polymorphism was detected in 19 (79.2%) of 24 patients at baseline, with mutation frequencies of 6.2–93%. The prevalence was much higher than 12% in Europe, as reported by previous studies. When the treatment outcome was considered, no difference was found between patients who achieved sustained virological response (SVR) and who didn’t (75% vs 81.3%, p = 1.00). Treatment of PegIFN plus RBV for 48 weeks did not change the frequency of this mutation. Interpretation: The NS5B substitution C316N associated with treatment failure was identified with a quite high prevalence in this study. The detection of this mutation should be considered for patients in Asia receiving treatment of sofosbuvir in the near future. http://dx.doi.org/10.1016/j.jcv.2015.06.039