Hepatitis B Virus DNA PCR Research Articles

Accuracy of transient elastography in Identifying fibrosis among patients with Chronic hepatitis b: a cross-sectional Observational study

Background: Hepatitis B virus (HBV) infection is a significant global public health problem, affecting millions of people worldwide. Liver biopsy is the gold standard for assessing fibrosis but is invasive and carries post-procedural risks. Noninvasive tests to evaluate liver fibrosis are needed, and transient elastography is one such method. This cross-sectional observational study aimed to assess the accuracy of transient elastography in identifying fibrosis among patients with chronic hepatitis B.
 Methods: The study was conducted at the Department of Medicine and Hepatology, Sir Salimullah Medical College Mitford Hospital, Dhaka. The study subjects were patients diagnosed with chronic hepatitis B infection, both outpatient and inpatient cases. Data were collected from 25 patients using convenient and purposive sampling techniques. Demographic data, clinical examination, and relevant investigations were recorded in a structured case report form. Data were processed, registered, edited, computerized, and analyzed.
 Results: The study included 25 patients with a mean age of 26.8±6.5 years, predominantly male (80%). All patients were HBsAg positive, 12.0% were HBeAg positive, and 56.0% were antiHBe positive. The mean HBV DNA PCR was 1516293.4±7475976.3 IU/ml. The mean ALT level was 45.4±13.8 U/L. The mean liver stiffness measurement (LSM) was 7.5±2.3 kPa, with 64.0% of patients showing fibrosis stage F0-F1. The mean periportal +/- bridging necrosis was 2.92±1.58, intralobular degeneration and focal were 1.16±0.55, portal inflammation was 2.52±0.87, histological activity index was 6.6±2.2, and fibrosis score was 1.44±1.04. Positive correlations were observed between LSM and histological activity index (r=0.239; p=0.251) and between LSM and fibrosis score (r=0.107; p=0.612).
 Conclusion: Our study demonstrates that transient elastography provides a non-invasive, easy, and cost-effective method for identifying fibrosis in chronic hepatitis B patients, serving as a potential alternative to liver biopsy.
 Bangladesh J Medicine 2023; 34(3): 199-205

S3028 Identifying and Screening At-Risk Patients for Hepatitis Delta Virus

Introduction: Hepatitis delta virus (HDV) is associated with the most severe forms of viral hepatitis with rapid progression to cirrhosis and hepatocellular carcinoma. The prevalence of HDV may be significantly higher than formerly acknowledged in the United States, making it doubly important to identify at-risk individuals. Case Description/Methods: A recently emigrated 29-year-old Afghani male with chronic hepatitis B virus (HBV) infection presented with abdominal pain, nausea and vomiting to an outside facility. He underwent a laparoscopic cholecystectomy for symptomatic cholelithiasis. He developed postoperative abdominal distension with imaging initially concerning for a large bile leak however ERCP and a diagnostic laparoscopy were unrevealing. Four liters of fluid were drained, and a surgical drain was placed which continued to drain up to two liters of fluid daily. Upon transfer to our hospital, diagnostic workup showed AST 583 U/L, ALT 452 U/L, alkaline phosphatase 88 U/L, total bilirubin 1.8 mg/dL, and albumin 2.8 g/dL. HBV DNA PCR was < 10 IU/mL. HBV core antibodies, HBV e antibody, and HBV surface antigen were reactive. HBV e antigen and HBV surface antibody were non-reactive. Ascites fluid studies showed a high serum ascites albumin gradient and low protein, prompting a liver biopsy that showed cirrhosis (image 1). The patient was ultimately found to be positive for HDV antibody, confirming a diagnosis of cirrhosis due to HDV superinfection. The patient had his drains removed and was discharged on oral diuretics and entecavir for treatment of HBV infection. Diuretic uptitration is ongoing with plans to enroll in a trial in the future pending clinical status. Discussion: In patients with chronic HBV who acquire HDV as a superinfection, 70-80% develop cirrhosis or hepatocellular carcinoma within 5-10 years. Superinfection is the likely cause of cirrhosis in our patient, as cirrhosis in a 29-year-old would not be expected with HBV infection alone. Early recognition of at-risk individuals can aid in faster diagnosis and earlier treatment. Screening should be considered in all individuals with a reactive HBV surface antigen, and especially in migrants from endemic areas, hemodialysis patients, healthcare employees, and IV drug users. Total HDV antibodies should be obtained for screening, with diagnosis confirmation by serum RT-PCR. Currently, therapeutic options are only available for compensated patients, with pegylated interferon alpha approved in the United States and buleviritide in Europe.Figure 1.: (A) Trichrome stain showing markedly active chronic hepatitis consistent with cirrhosis. (B) Immunohistochemical stains demonstrate patchy staining with HBV surface antigen but (C) negative for HBV core antigen.

S1198 Treatment and Hepatocellular Carcinoma Screening Patterns Among Black Patients With Chronic Hepatitis B

Introduction: Significant racial disparities exist in Hepatitis B virus (HBV) infection across the nation, from screening variability to treatment patterns and clinical outcomes. The Black community bears a disproportionately large disease burden. In this study, we sought to compare the disease courses between Black patients seen at a private tertiary referral center vs. those evaluated at a high-volume county safety net hospital. Methods: Using informatics, we identified adult patients with chronic HBV based on ICD-10 codes, who had ≥1 clinic visit between January 1, 2010 and December 31, 2020 at University of Miami Hospital (UM) and Jackson Memorial Hospital (JM). We conducted retrospective chart review to gather demographic and clinical data. We used descriptive statistics, Kruskal-Wallis, and Pearson’s chi squared tests to evaluate for differences between Black patients at each hospital with the significance interval set to p < 0.05. Analyses were conducted using STATA 17.0. Results: Of 649 Black patients detected by informatics, 364 had confirmed HBV with 306 seen at UM and 58 at JM. Compared to JM, UM patients were significantly older, English speakers, and privately insured (p < 0.001) (Table). Most common birth countries included Haiti and the USA, among others (Figure). 78.8% of UM patients were evaluated by Hepatology, compared to 41.1% at JM. More JM patients had active hepatitis, and 15.5% had a high viral load (HBV DNA PCR >20,000) compared to 3.3% at UM. Patients seen at UM were more likely to be treated, 52.9%, vs. 24.6% at JM and placed on newer regimens (ie Tenofovir [TAF] and Entecavir). UM patients were significantly more likely to undergo HbeAg and HbeAb testing, compared to their JM counterparts. Black patients at JM were significantly more likely to have alcoholic liver disease, ascites, and orthotopic liver transplantation. Although more UM patients underwent surveillance for hepatocellular carcinoma (HCC), there was ultimately no significant difference in patients being diagnosed with HCC between hospitals. Conclusion: These findings suggest that significant differences in demographics, referral patterns, laboratory evaluation, and management exist between Black patients at UM vs. JM, despite both hospitals being affiliated with the same academic institution. Our analysis on JM patients is ongoing and will be crucial to inform future interventions aimed at standardizing and providing evidence-based HBV care across medical centers.Figure 1.: Geographic distribution of Black patients’ most common birth countries. Green UM, Orange JM. Table 1. - Comparison of demographics, screening, and management patterns between Black patients at JM vs. those at UM Variable Blacks @ JM (n = 58) Blacks @ UM (n = 306) p-value Median Age, years (IQR) 54 (45-63) 59 (47-66) 0.0001 Male Gender, % 68.3 63.7 0.24 US Born, % 50.9 32.7 0.012 Department seen by, %Hepatology 41.4 78.8 < 0.001 General Internal MedGastroenterology 62.11.7 8.814.1 < 0.0010.008 HbeAg / HbeAb checked, % 52.6 78.5 < 0.001 Active hepatitis, % 19 2.9 < 0.001 High viral load ( >20,000), % 15.5 3.3 < 0.001 Currently on Treatment, % 24.6 52.9 < 0.001 Tenofovir TDF TAF EntecavirAdefovirLamivudine 13.813.713.113.40.68.6 31.412.11.75.201.6 0.0070.740.00140.0780.540.003 Alcoholic Liver Disease, % 14 1.8 < 0.001 Ascites, % 22.8 7.3 < 0.001 OLT, % 26.3 1.8 < 0.001 HCC Screening, %FibroscanAFPSurveillance imaging (< 6 months) 17.542.1158.9 3178.377.9 0.04< 0.0010.003 Diagnosed with HCC 5.1 5.9 0.71 AFP = alpha-fetoprotein; OLT = orthotopic liver transplantation; HCC = hepatocellular carcinoma.

Hepatitis B Seroprevalence among Egyptian University students in the postinfant compulsory vaccination period

Background The three doses constituting the hepatitis B (HB) vaccine series administered intramuscularly at birth, 1 month, and 6 months induce a protective antibody response (anti-HBs &gt;10 IU/l) in more than 90% of recipients. Aim To analyze the long-term immunity and effectiveness of hepatitis B virus (HBV) vaccination and to detect hepatitis B infection situation and its risk factors among an adequate number of the university students in the postcompulsory infant vaccination period. Patients and methods A total of 400 university students (aged 17–25 years) were screened for quantitative detection of hepatitis B surface antibody, hepatitis B surface antigen (HBs Ag), and total hepatitis B core antibody (HBc Ab) using commercially available kits. HBV DNA PCR was evaluated in repeatedly positive HBs Ag and/or total HBc Ab. Results Nonprotective titer less than 10 IU/l was detected in 218/400 (54.5%), whereas protective titer more than or equal to 10 IU/l was detected in 182/400 (45.5%). Overall, 29/400 (7.3%) were positive for HBs Ag and 50/400 (12.5%) were positive for total HBc Ab, whereas only six (1.5%) were positive for HBV DNA PCR. Moreover, 45/400 (11.4%) students had a history of HBV vaccine booster dose in the protective group. Conclusions Marked reduction in HBs Ab titer was observed among university students where nonprotective titer less than 10 IU/l was detected in 54.5%. Chronic HBV inactive carrier was detected in 5.3%, chronic hepatitis B was detected in 1.5%, and resolved HBV infection was detected in 11%.

Significance of Hepatitis B Virus Diagnosis by Real-time Polymerase Chain Reaction over Serological Markers in Hepatitis B Virus Patients

Hepatitis B virus (HBV) is the leading cause of viral hepatitis, as currently over 2 billion people have HBV infection worldwide. Nucleic acid assay and quantitative hepatitis B surface antigen (HBsAg) have been developed for diagnostic and therapeutic monitoring of patients with HBV infection. These tests might also show correlation between HBV DNA and HBs serostatus. The study aimed to find and analyze the frequency and impact of HBsAg seropositivity among patients revealed HBV DNA negative level through quantitative estimation of both seromarkers. Real-time polymerase chain reaction (RT-PCR) and Elecsys assays were used for quantitative estimation of HBV DNA and HBs antigen, respectively. A total of 256 blood samples were used from patients referred for either diagnostic purpose and/or HBV viral load monitoring after antiviral therapy. Blood profile analysis showed 12.26% HBs antigen seropositivity among patients revealed negative for nucleic acid assay for HBV DNA. Positive HBs antigen titers ranged from 1000–50,000 COI, with seronegative anti-HBs antibody test for all samples tested positive for HBs antigen. This study delineated that negative or undetectable quantitation of HBV DNA level does not exclude HBV infection; as the level might fluctuate in different phases of HBV replication. This gives an impression and raising a question about significance of replacing test for HBsAg with quantitation of HBV DNA PCR assay. Thus, the study refers to a special HBV profile outside the classical pattern.

Occult Hepatitis B in Patients Undergoing Chemotherapy

Introduction: The hepatitis B virus (HBV) is one health problem in Iran. Occult hepatitis B (OBI) is diagnosed by the detection of HBV DNA in the serum or liver tissue of patients for whom other serology, especially HBsAg and HBcAg, are negative. The current study aimed to determine the prevalence of OBI in patients who refer for chemotherapy. Methods: All patients referring for chemotherapy to the oncology clinic of Baqiyatallah Hospital in Tehran, Iran from 2012 to 2013 were selected as the population of this cross-sectional study. Samples of 5 mL of blood were taken from each subject and assayed for HBsAg and HBcAb. If HBV markers were negative, a qualitative HBV DNA PCR was done to detect OBI. The data was analyzed using SPSS 17 software, and the frequency test was used to determine prevalence. Results: In this study, 251 candidates for chemotherapy were recruited. Of this number, 114 (45%) patients were women. Mean patient age was 52.6 years. A total of 149 (59%) study subjects were employed, and the others were housekeepers or self-employed. About 67 (26.6%) patients had a history of one dose and 40 (15.9%) patients had a history of complete HBV vaccination. HBsAg was positive in 6 (2.4%) and HBcAb was positive in 33 (13.1%) patients; qualatative HBV DNA PCR was positive in 2 (0.8%) patients. Conclusion: This study revealed that OBI infection has a low prevalence, but patients referring for chemotherapy are at risk for its reactivation. On the other hand, isolated HBcAb is also a risk factor; therefore, in addition to HBsAg, it is recommended that HBcAb be evaluated before chemotherapy.

Inclusion of Seromarkers to Exclude Hepatitis B Virus Infection via Transfusion in Voluntary Blood Donors

Introduction: Hepatitis B Virus (HBV) infection is a serious viral infection causes acute and chronic hepatitis. Transfusion of blood and blood products is the predominant route of transmission of HBV. As per NACO guidelines, in India, donor blood is screened for HBsAg, anti-HCV antibodies, anti-HIV antibodies; slide/card rapid screening for malarial parasites and VDRL for Syphilis.Blood banks in India are only equipped to carry out rapid card test or ELISA for routine screening of HBV, HCV and HIV infections. In developed countries use Nucleic Acid Testing (NAT) were in place for screening and diagnosis of viral infections during window period. NAT assays are highly sensitive, specific and robust but require skilled manpower and specialized infrastructure and thus may not be cost-effective.Objectives: To explore the current sero-prevalence of HBsAg, Anti-HCV antibodies and anti-HIV antibodies (and other relevant seromarkers) by routine screening tests in a cohort of voluntary blood donors and to determine the efficiency of NAT over the routinely used rapid card and ELISA tests. To include any additional seromarker of HBV infection to detect the antigen in the window period.Methods: In this cross-sectional observational study, 3160 voluntary blood donors were screened for HBsAg. Samples were also subjected to screening of additional seromarkers such as HbeAg, anti-HBs and anti-HBc antibodies. Thirty samples were randomly selected from each HBsAg positive and HBsAg negative groups and were subjected to HBV DNA PCR for viral load estimation.Results: ELISA testing is more specific than rapid card test in detecting HBV infection, whereas NAT testing was beneficial in identification of added HBV infection. Both ELISA and rapid card test methods were found to be equally sensitive and specific in diagnosing HCV and HIV infections. NAT testing did not yield any significant increase in positive results compared to serological/rapid methods. HBeAg was found to be positive in 2 samples which are tested negative for HBsAg. HBV DNA was also found to be positive in one sample which is negative for HBsAg (1 out of 30), which might be cryptic HBV infection.Conclusion: Though NAT would be the choice of screening for Hepatitis B, it not cost-effective. Therefore, inclusion of one or more seromarkers for screening HBV, such as HBeAg, might be helpful in preventing transfusion transmitted infection.

Performance of two real-time PCR assays for hepatitis B virus DNA detection and quantitation

In-house developed real-time PCR (qPCR) techniques could be useful conjunctives to the management of hepatitis B virus (HBV) infection in resource-limited settings with high prevalence. Two qPCR assays (qPCR1 and qPCR2), based on primers/probes targeting conserved regions of the X and S genes of HBV respectively, were evaluated using clinical samples of varying HBV genotypes, and compared to the commercial Roche Cobas AmpliPrep/Cobas TaqMan HBV Test v2.0. The lower detection limit (LDL) was established at 104 IU/ml for qPCR1, and 91 IU/ml for qPCR2. Good agreement and correlation were obtained between the Roche assay and both qPCR assays (r = 0.834 for qPCR1; and r = 0.870 for qPCR2). Differences in HBV DNA load of > 0.5 Log10 IU/ml between the Roche and the qPCR assays were found in 49/122 samples of qPCR1, and 35/122 samples of qPCR2. qPCR1 tended to underestimate HBV DNA quantity in samples with a low viral load and overestimate HBV DNA concentration in samples with a high viral load when compared to the Roche test.Both molecular tools that were developed, used on an open real-time PCR system, were reliable for HBV DNA detection and quantitation. The qPCR2 performed better than the qPCR1 and had the additional advantage of various HBV genotype detection and quantitation. This low cost quantitative HBV DNA PCR assay may be an alternative solution when implementing national programmes to diagnose, monitor and treat HBV infection in low- to middle-income countries where testing for HBV DNA is not available in governmental health programmes.

Interleukin-17-producing CD4+T cells in patients with chronic hepatitis B

Objective Our aim is to evaluate the expression of interleukin-17-producing CD4 + T cells (Th17) in peripheral blood of patients with chronic hepatitis B (CHB) and whether these cells show a potential to exacerbate liver damage during chronic hepatitis B virus (HBV) infection. Background Interleukin-17-producing CD4 + T cells (Th17)-mediated immune response has been shown to play a critical role in inflammation-associated disease; however, its role in chronic HBV infection remains unknown. Here, we characterized peripheral Th17 cells and analyzed their association with liver injury in HBV-infected patients. Participants and methods This study included 15 patients with CHB without cirrhosis (11 men and four women), 15 patients with CHB with cirrhosis (eight men and seven women), and 15 healthy controls (10 men and five women). Laboratory investigations including complete blood picture, liver function tests, hepatitis viral markers (HBsAg, anti-HCV-Ab), a-fetoprotein, and HBV DNA PCR were performed for all participants. Evaluation of the level of Th17 cells in the peripheral blood was carried out using the flow cytometry technique. Results The data showed that the frequency of circulating Th17 cells was increased in patients with CHB viral infection compared with the healthy controls, and was significantly higher in patients with CHB-associated cirrhosis. In addition, the increasing levels of circulating Th17 cells were correlated positively with serum alanine transaminase levels. Conclusion Our results showed that Th17 cells were highly enriched in the peripheral blood of CHB patients, and their levels were related to disease progression, suggesting its role in exacerbation of liver damage during chronic HBV infection.

Highly Sensitive Detection of Hepatitis B Virus Surface Antigen by Use of a Semiautomated Immune Complex Transfer Chemiluminescence Enzyme Immunoassay

The performance of hepatitis B surface antigen (HBsAg) screening assays is continuously improved to reduce the risk of transfusion-associated hepatitis B. In this study, a semiautomated immune complex transfer chemiluminescence enzyme immunoassay (ICT-CLEIA) for the detection of HBsAg, which is as sensitive as hepatitis B virus (HBV) DNA PCR, was developed; the ICT-CLEIA assay performance was compared with the performance of the Architect HBsAg QT assay and HBV DNA PCR. The specificities in the initial assay and after retesting were 99.50% (1,988/1,998 samples) and 99.95% (1,997/1,998 samples), respectively. The analytical detection limit was determined to be 0.2 mIU/ml using the 2nd International WHO HBsAg standard, and the cutoff value (0.5 mIU/ml) of the ICT-CLEIA assay was 8.0 standard deviations (SD) above the mean of the HBsAg-negative specimens. The ICT-CLEIA assay could detect HBsAg even in the presence of anti-HBs antibodies and demonstrated a 23.6-day-shorter window period using commercially available HBsAg seroconversion panels than the Architect HBsAg QT assay. Furthermore, the monitoring of the viral kinetics by the ICT-CLEIA assay and the HBV DNA PCR produced very similarly shaped curves during both the HBsAg seroconversion and reverse seroconversion periods. Therefore, the ICT-CLEIA assay may be useful not only for an earlier detection of HBV reactivation but also for the monitoring of hepatitis B patients.

Sa1019 Incidence of Hepatitis B Surface Antigen and Hepatitis B Core Antibody in a Screening Population Undergoing Chemotherapy

Background: Reactivation of hepatitis B virus (HBV) infection is a well-recognized complication after immunosuppression from chemotherapeutic agents and can lead to significant morbidity and mortality. We previously reported 23 HBV reactivations in patients of diverse nationalities without association with a particular malignancy or medication. Our institution subsequently initiated a protocol in which all new patients receiving immunosuppressive therapy are screened for HBV and offered anti-viral prophylaxis. The aim of this study is to report the prevalence of HBV surface antigen (HBsAg) and HBV core antibody (HBcAb) positivity since the inception of this screening program.Methods:We conducted a prospective study of all patients at Memorial Sloan-Kettering Cancer Center who were started on immunosuppressive therapy and screened for HBV from May 2009 to November 1, 2012. Patients were screened using tests for HBsAg and HBcAb. If either of these were positive, PCR for HBV DNA was reflexively measured. Patient demographics --including type of malignancy-were recorded. Results: Between May 1 2009 and November 1 2012, 17183 patients met criteria for screening and 12,328 patients (median age 64 years [24-95], 54.4% male) were screened for HBV prior to initiation of immunosuppression (71.75% compliance). Testing revealed 78(0.6%) patients positive for both HbsAg and HbcAb (median age 58, 69.2% male).Of this group, 47/78 (60.26%) had detectable HBV DNA PCR (955766.7 mean, 521 median) .Primary diagnoses of these patients included lymphoma 6.4%, leukemia 2.6%, and myriad solid tumors 91.0%. In addition, 994 patients (8.1%) were negative for HBsAg and positive for HBcAb (median age 58, 53.2% male), Diagnoses among this group included 6.0% lymphoma, 2.0% leukemia and 92.0 % solid tumor. Of this group, 7 (0.7%) had detectable HBV DNA PCR ([41-1,740,000 IU/mL]). Conclusion: In our population screened for HBV prior to initiation of immunosuppression, we report a prevalence of 0.6% for HBsAg positive patients and 8.1% for HBsAg negative, HBcAb positive patients. Screening of these

Hepatitis B reactivation after therapy for non-Hodgkin lymphoma: A case report with review of literature

The natural course of hepatitis B virus (HBV) infection depends on the immune status of the host. In cancer patients, as the consequence of immune suppression due to chemotherapy and malignant disease itself, the balance between replicative potential of the virus and immune response of the host is disrupted leading to acute HBV infection or reactivation. We present a case of HBsAg positive, diffuse large B cell gastric lymphoma patient CD20+ staged IB, treated with six cycles of R-CHOP protocol and two cycles with rituximab monotherapy. Five months after the successful anticancer treatment, patient developed reactivation of chronic HBV infection (ten-fold increase in liver enzymes, HBsAg+, IgM antiHBc+, HBeAg(-), and HBV DNA 5?106 copies/ml). Antiviral therapy with lamivudine was started. Four weeks after the antiviral therapy initiation liver enzymes were in normal ranges. One year after the start of antiviral treatment HBV DNA PCR test did not detect any viral particles. The patient is in complete remission of malignant disease, and still receiving therapy with lamivudine. HBV screening in cancer patients is necessary in order to provide a prompt antiviral therapy and to prevent postponement or even cessation of planned anticancer treatment. HBsAg positive patients should start prophylactic antiviral treatment before the start of immunosuppressive treatment. Chemotherapy protocols consisting rituximab and corticosteroids significantly increase the risk of reactivation. If reactivation is diagnosed in course of chemotherapy, the therapy should be stopped and antiviral treatment should be applied as soon as possible. Treatment with lamivudine is continued at least 6 months after the chemotherapy end.

Elecsys Hepatitis B Surface Antigen Quantitative Assay: Performance Evaluation and Correlation with Hepatitis B Virus DNA during 96 Weeks of Follow-up in Chronic Hepatitis B Patients

BackgroundTreatment for chronic hepatitis B aims to suppress virus replication and virus sequestration in hepatocytes. Covalently closed circular (ccc) DNA is the template for transcription of viral genes and is responsible for viral persistence. However, limited data are available for quantification of hepatitis B surface antigen (HBsAg) in Korea.MethodsWe evaluated the Elecsys HBsAg II quant assay (Roche Diagnostics, USA) for within-run, between-run, and between-day precisions, linearity, carryover, and clinical specificity. In total, 156 serum samples were evaluated for correlation between HBsAg and hepatitis B virus (HBV) DNA. Serial samples were obtained from 10 patients at 0, 12, 24, 48, 72, and 96 weeks during follow-up.ResultsThe assay detected HBsAg in a linear range of 0.5-48,696 IU/mL. Within-run, between-run, and between-day CVs were 2.9-4.1%, 0-1.5%, and 1.5-4.9%, respectively. Cross-reactivity between potentially interfering substances was absent, and the carryover rate was 0.00002%. The correlation of measurements between the Elecsys assay and HBV DNA PCR was weak (r=0.438, P=0.002). For predicting virologic response, cutoff values of 10,275 IU/mL and 3,846 IU/mL at 12 and 24 weeks after treatment initiation showed positive predictive values of 77.1% and 85% and negative predictive values of 84.6% and 50%, respectively.ConclusionsThe Elecsys HBsAg II quant assay showed good performance for precision, linearity, carryover rate, and specificity. HBsAg level at baseline, 12 weeks, and 24 weeks after treatment initiation can predict virologic response, and the assay can be used for HBsAg quantification in clinical practice.

Sa1045 Factors That Affect the Improvement of Liver Stiffness in Patients With Chronic Hepatitis B

Background and aims Long term suppression of Hepatitis B virus (HBV) replication with nucleos(t)ide analogues (NA) is associated with biochemical improvement and viral suppression. However the factors affecting this response remain unclear. Method A cohort of 107 HBV infected patients undergoing treatment with NA at Royal North Shore Hospital (20002011) was studied. Demographic, viral, biochemical and histological data were collected prospectively during treatment and subjected to univariate and multivariate analysis. Result Seventy-eight percent of the total cohort were Asian, 70% were males and 47% had advanced fibrosis (F≥3) at baseline. Mean HBV DNA PCR was 5.62 log IU/mL and mean ALT was 129 IU/L at baseline. Mean duration of treatment was 66 months. There were 42 eAg positive and 65 eAg negative patients. In the combined cohort, 64% had undetectable DNA at week 48. Negative eAg at baseline, lower DNA at baseline, no interferon use (before or during the therapy), normalised ALT at week 48 and older age were associated with undetectable DNA at week 48. (p<0.05) However, logistic regression analysis identified baseline eAg negativity as the only significant predictor of undetectable DNA at week 48. (p=0.02) Sixtytwo percent had normalised ALT at week 48, and associated factors were undetectable DNA at week 48, shorter time to undetectable DNA and higher ALT at baseline. (p<0.05) Logistic regression identified shorter time to lowest DNA as the only predictor of normalised ALT at week 48. (p=0.02) In patients with positive eAg at baseline, 55% and 7% achieved eAg seroconversion (or loss) and sAg seroconversion (or loss) respectively. Older age and advanced fibrosis were significantly associated with eAg seroconversion and/or eAg loss. (p≤0.05) No patients developed progressive liver failure while on treatment in the absence of hepatocellular cancer. Conclusion NA therapy in the clinic setting is associated with effective long term viral suppression with improved eAg seroconversion in older patients and those with advanced fibrosis. Higher rates of sAg seroconversion or loss were observed in patients with positive eAg at baseline.

True prevalence of twin HDV-HBV infection in Pakistan: a molecular approach

Hepatitis Delta Virus (HDV) infects only patients that are already infected by hepatitis B virus (HBV) because this is sub satellite virus which depends on and propagate only in the presence of HBV. HDV causes co-infection or super infection with sever complication as compared to only HBV infection. No study on molecular level on HDV is available from this region; therefore, the aim of this study was to found out the molecular epidemiology of HDV (as a co-infection with HBV) in different geographical regions of Pakistan.Total 228 HBsAg positive samples were received for the study from different geographical regions of the country. Only HBV DNA PCR positive samples were further utilized for the presence of HDV RNA. For this purpose, HDV RNA and HBV DNA was extracted and amplified using reverse transcriptase polymerase chain reaction (RT-PCR), nested PCR and real-time PCR.Out of the total 228 HBsAg positive samples, HBV DNA was detected in total 190 (83.3%) samples belonged to different patients. Of these 190 patients, HDV RNA was observed in 53 (28%) patients. Of the 53 HDV positive cases, 37 (69.8%) were males and 16 (30.2%) were female patients. The percentage of dual infection was found higher significantly (p < 0.05) in male patients as compared to female patients. Total 41 (26.8%) patients were below 40 years and 13 (31.7%) were above 40 years of age. No significant difference was seen in patients with ages above or below 40 years. In the provinces of Sindh, Khyber Pakhtoonkhaw and Punjab the observed prevalence of HDV was 67%, 6% and 4% respectively.In conclusion, the HDV infection is not uncommon in Pakistan and its prevalence is higher significantly in the Province of Sindh (p < 0.01) and male six (p < 0.05).

Virological and clinical characteristics of hepatitis delta virus in South Asia.

Background & AimsThere is a paucity of data on the impact of hepatitis D virus (HDV) in patients with hepatitis B virus (HBV) infection from South Asia. We studied the impact of HDV co-infection on virological and clinical characteristics.MethodsWe collected data of 480 patients with HBsAg positive and a detectable HBV DNA PCR, who presented to the Aga Khan University, Karachi and Isra University in Hyderabad, Pakistan in the last 5 years. HDV co-infection was diagnosed on the basis of anti-HDV. ALT, HBeAg, HBeAb and HBV DNA PCR quantitative levels were checked in all patients. We divided all patients into two groups based on anti-HDV, and compared their biochemical, serological & virological labs and clinical spectrum. Clinical spectrum of disease included asymptomatic carrier (AC), chronic active hepatitis (CAH), immuno-tolerant phase (IP), and compensated cirrhosis (CC).ResultsHDV co-infection was found in 169 (35.2%). There were 164 (34.6%) HBeAg positive and 316 (65.4%) HBeAg negative patients. Mean ALT level was 66 ± 73 IU. 233 (48.5%) had raised ALT. HBV DNA level was ≥ 10e5 in 103(21.5%) patients. Overall, among HBV/HDV co-infection, 146/169 (86.4%) had suppressed HBV DNA PCR as compared to 231/311 (74.3%) patients with HBV mono-infection; p-value = 0.002. Among HBeAg negative patients 71/128(55.5%) had raised ALT levels among HBV/HDV co-infection as compared to 71/188 (37.8%) with HBV mono-infection (p-value = 0.002); levels of HBV DNA were equal in two groups; there were 27/128 (21%) patients with CC among HBV/HDV co-infection as compared to 23 (12%) in HBV mono-infection (p-value = 0.009); there were less AC (p-value = 0.009) and more CAH (p-value = 0.009) among HBV/HDV co-infection patients. Among HBeAg positive patients, serum ALT, HBV DNA levels and the spectrum of HBV were similar in the two groups.ConclusionsHBV/HDV co-infection results in the suppression of HBV DNA. A fair proportion of HBV/HDV co-infected patients with HBeAg negative have active hepatitis B infection and cirrhosis as compared to those with mono-infection.

A Study on HBV Infection in HBV-HIV Co-infected Patients from Eastern India

© 2011, INASL 3 negative samples. For HBV DNA positive cases, HBeAg status was checked and direct sequencing of surface gene, BCP and PC region were done. Surface gene sequences were used to confirm the HBV genotypes/subgenotypes by phylogenetic analysis. The viral load was measured by real time PCR. Results: After initial screening, 204 samples were found to be anti-HBc positive but HBsAg negative. From these 204 samples 31 (15.2%) were found to be positive for HBV DNA. HBV genotype A was present in 9.7% and rest of the samples were HBV/D (90.3). Three subgenotypes of HBV/D was identified, viz D2 (35.7%), D3 (50.0%) and D5 (14.3). In the BCP region most common mutations were C1752, C1753 and triple mutation C1753/T1762/A1764. In 48.4% cases the presence of C1773 was noticed. Notably all the samples were found to be HBeAg negative. No mutation was found in the PC region. Conclusion: Interestingly higher rate of variability in the BCP region, especially C1752, C1753/T1762/A1764 triple mutations and C1773 was noticed. These carriers carrying the mutant strains should be followed up at regular intervals to assess the clinical progression. Conflict of Interest: None A Study on HBV Infection in HBV-HIV Co-infected Patients from Eastern India A Pal*, M Bandopadhyay*, A Biswas*, R Panigrahi*, SK Guha**, S Chakrabarti†, R Chakravarty* *ICMR Virus Unit, Kolkata **Department of Medicine, School of Tropical Medicine, Kolkata †National Institute of Cholera and Enteric Diseases, Kolkata Background: Hepatitis B virus (HBV) and HIV co-infection is frequent due to shared routes of transmission. With improved control of HIV disease with HAART, liver disease has emerged as one of the leading causes of deaths in HIV patients. It has been reported that co-infection with HIV complicates the natural history, diagnosis and management of HBV infection. In India, the study regarding HBV-HIV co-infection is sparse although the prevalence of both viruses is considerable. Objective: The study is aimed to characterize HBV infection in hepatitis B surface antigen (HBsAg) positive cases of HBV-HIV co-infected patients. Methods: Total 48 HIV patients who were registered in our unit for HBV screening were included. Serological markers of HBV and HIV were tested using enzyme linked immunoassays. Detection of HBV DNA was done by nested PCR, followed by genotyping using RFLP. Result: Thirty-two HIV patients were detected positive for HBsAg and HBV DNA resulting in 66.67% of selected HIV patients to be co-infected with HBV. 75% of 32 co-infected patients were also positive for HBeAg. Sexual transmission (47.62%) was found to be more associated with HBV transmission than blood transfusion and surgical operation (9.52% in both cases). When HBV genotype distribution was correlated with CD4 cell count, three genotypes [HBV/A (21.34%), HBV/C (7.14%) and HBV/D (64.29%)] were found to be related with CD4 cell count 6 months included. Patients with cirrhosis, hepatocellular carcinoma, antiviral treatment, HIV/HCV coinfection, excluded. Detailed clinical examination, LFT, USG abdomen, AFP, HBsAg ELISA titers, HBeAg and HBV DNA PCR (Roche Amplicor) assays done. OGD done when indicated. Anti-HBe antibody assessed in HBeAg negative patients. Pearson’s correlation coefficient (r) used for correlation between variables, analyzed using SPSS. 03_JCEH-Abstract.indd 3 3/18/2011 11:13:03 AM

Probing rate of YMDD motif mutant in lamivudine treatment of Iranian patients with chronic hepatitis B virus infection

Background:Lamivudine is an analog nucleoside used for the treatment of chronic hepatitis B virus (CHV) infection. Studies showed that prolonged therapy could induce lamivudine resistance hepatitis B virus (HBV) variants (YMDD motif). In this study, the rate of YMDD motif mutants is determined in lamivudine-treated CHV patients in Iran.Materials and Methods:Thirty-three serum specimens of patients with chronic hepatitis who had been treated with lamivudine were included in the study. Serum samples of patients were tested by PCR flash and RFLP as well as tested for HBeAg, HBsAg, and liver enzymes (ALT and AST).Results:Out of the 33 patients enrolled in this study, 82% were male and 18% female, respectively. Mean ALT levels were between 20 and 100 IU/1. HBeAg was positive in 76% of the patients whereas HBs was positive only in 61% of the patients. Furthermore, in 28 patients liver biopsy grade was between 2 and 17 having the stage of 1–6. Moreover, HBsAg negative and HBeAg positive were observed in 30% of the patients.Conclusion:During therapy, it was found that patients with lamivudine incidence YMDD mutation were approximately 14%. The ALT levels were also reduced in these patients. This study revealed that there was a significant difference between HBeAg, grade, and YMDD mutation whereas no significant different was observed between HBsAg and HBV DNA PCR. Conclusively, it was found that a significant difference exists between YMDD mutation and lamivudine therapy (17% of patients were HBsAg negative and PCR positive).

Reactivation of chronic hepatitis B virus in cancer patients receiving immunosuppression: The case for screening.

9088 Background: Reactivation of hepatitis B virus (HBV) infection following immunosuppressive therapy can cause significant morbidity and mortality. This study aims to estimate the frequency and clinical consequences of reactivation of HBV in a diverse group of cancer patients (pts) receiving immunosuppression. Methods: We retrospectively reviewed all pts at Memorial Sloan-Kettering Cancer Center between 2003 and 2009 who had an HBV DNA PCR>1000 to identify cases of reactivation of HBV. The medical record was used to distinguish between reactivation and de novo infection. Pt demographics, type of malignancy, immunosuppressive agent, and clinical outcome were recorded. Results: Between 2003 and 2009, 241 pts had HBV PCR>1000 copies including 23 with associated abrupt increase in serum ALT. Twenty-two of these pts (median age: 53; 45% Asian descent) had no risk factors for acute HBV, suggesting reactivation. Immunosuppressive therapy began a median of 21 days (range 1-175) prior to reactivation. Varied chemotherapeutic agents were used; 1 pt received high dose steroids only. Seven (32%) patients had lymphoma, and 15 (68%) had other tumors: hepatocellular carcinoma (2), glioblastoma (2), leukemia (2), multiple myeloma (1), esophageal ca (1), sarcoma (1), breast ca (1), lung ca (1), jejunal ca (1), pancreatic ca (1), ovarian ca (1), and neuroendocrine tumor (1). Four (21%) pts died from liver failure; 19 (86%) required hospitalization (median LOS 5 days (range 2-33)). Four pts had clinically significant delays in cancer treatment due to HBV reactivation. Three pts were transferred for liver transplant evaluation; 1 pt underwent transplant. Conclusions: Reactivation of HBV during chemotherapy-induced immunosuppression is a completely preventable disease with substantial morbidity and mortality. Our 3 cases per year of clinically significant HBV reactivation occurred in patients of various nationalities without association with a particular malignancy or medication. Because of these findings, we now screen all patients initiating immunosuppressive therapy for HBV; subsequent use of antiviral prophylaxis is based on planned treatment and HBV risk group. No significant financial relationships to disclose.

Comparison of Clinical Application of the Abbott HBV PCR Kit and the Versant HBV DNA 3.0 Test to Measure Serum Hepatitis B Virus DNA in Taiwanese Patients

With an estimated 350-400 million people worldwide chronically infected with hepatitis B virus (HBV), and the subsequent serious complications caused by liver damage including cirrhosis, liver failure, and hepatocellular carcinoma, HBV infection remains a global health issue, particularly in Taiwan, an HBV-hyperendemic area. Sensitive and accurate quantification of HBV DNA is necessary to monitor patients with chronic hepatitis B who are receiving antiviral therapy to determine treatment response and adapt therapy. We evaluated and compared the clinical performance of two HBV DNA assays based on different technologies: the RealArt HBV PCR Kit (Abbott HBV DNA PCR kit, real-time polymerase chain reaction assay, detection limit: 27 IU/mL) and the VERSANT bDNA 3.0 assay (Bayer, branched DNA signal amplification assay, detection limit: 357 IU/mL). Serum levels of HBV DNA in 173 chronic HBV carriers were determined using both the RealArt HBV PCR Kit and the VERSANT bDNA 3.0 test. Of the 173 samples analyzed for baseline viral load detection, HBV DNA was quantifiable in 147 patients (82.1%) by the RealArt HBV PCR Kit, which was significantly higher than the 92 (53.2%) samples quantified by the VERSANT bDNA 3.0 assay. A total of 86 (49.7%) samples were quantifiable by both assays, whereas 25 (14.5%) were below the detection limit of both assays. The HBV DNA quantification values measured by the RealArt HBV PCR Kit and the VERSANT bDNA 3.0 assay were positively correlated (Spearman's rank correlation coefficient r = 0.932, p < 0.001). On average, the results derived from the RealArt HBV PCR Kit were 0.67 log lower than those of the VERSANT bDNA 3.0 assay. HBV DNA concentrations were significantly higher in 63 HBV e antigen (HBeAg)-seropositive patients than in 110 HBeAg-seronegative patients (5.42 +/- 2.34 logs vs. 3.21 +/- 2.27 logs, p < 0.001). The RealArt HBV PCR Kit is more sensitive and has a wider dynamic range than the VERSANT bDNA 3.0 assay in the clinical setting of chronic hepatitis B patients. The sensitivity and wide dynamic range of the PCR assay allow optimal monitoring and timely adaptation of antiviral therapy. Nevertheless, the HBV DNA values measured by the RealArt HBV PCR Kit and the VERSANT bDNA 3.0 assay were significantly correlated.