Efficacy of accelerated hepatitis B vaccination program in patients being actively treated for hematologic malignancies.

BACKGROUND: The goal of this study was to conduct an accelerated vaccination program and to determine its efficacy in patients susceptible to hepatitis B virus (HBV) receiving chemotherapy because of their hematologic malignancies. METHODS: Over a one-year period, a total of 327 patients who were diagnosed as having a hematologic malignancy were serologically analyzed in terms of HBV infection. Of those found to be susceptible to HBV infection, a total of 42 patients consisting of 16 females and 26 males were enrolled in the accelerated vaccination program. All the patients were administered a 20-microg yeast-derived recombinant hepatitis B vaccine on days 0, 14, and 28. Anti-HBs titers above 10IU/l at 1 and 3 months after the final dose were accepted as protective. RESULTS: A total of 146 (44.6%) patients were susceptible to HBV, while 13 (4.0%) were carriers, 28 (8.6%) were vaccinated, and 113 (34.5%) had had a previous HBV infection. A total of 42 patients (16 females and 26 males, mean age 34.5+/-10.9 years) were enrolled in the vaccination program. Overall, 23.8% (10/42) of the patients in the program had developed anti-HBs at one month after the last vaccination. CONCLUSIONS: Poor results obtained by different vaccination programs suggest the need for alternative strategies to prevent the disease.

Restriction fragment length polymorphism analysis and direct sequencing for determination of HBV genotypes in a Turkish population.

The aim of this study was to analyze the restriction fragment length polymorphism and direct sequencing results in genotyping of hepatitis B virus from a Turkish population in a clinical virology laboratory. Serum samples of 54 chronic hepatitis B patients attending the Ege University Hospital were studied. Sequences of partial S gene PCR products were analysed and RFLP was performed. Fifty-three isolates could be identified by direct sequencing as genotype D. One sample needed to be cloned and determined as genotype D. Forty-two isolates were genotyped as D with RFLP according to published determinative patterns. Twelve isolates had undefined patterns. Eight of them suggested a mixture of isolates with different patterns and cloning of these samples confirmed the presence of heterogeneous isolates. Four isolates with undefined pattern were determined as genotype D by direct sequencing. All the studied isolates were genotype D. The results of this studied population suggest that RFLP is suitable for HBV genotyping in a routine clinical virology laboratory setting. However sequence analysis and even cloning may be needed to clarify indeterminate results.

Hepatitis delta virus (HDV) genotypes in patients with chronic hepatitis: molecular epidemiology of HDV in Turkey.

OBJECTIVE: Analysis of hepatitis delta virus (HDV) isolates from around the world has indicated that there are at least three phylogenetically distinct genotypes with different geographic distributions. The aim of this study was to determine the distribution of HDV genotypes by direct sequencing in patients with chronic delta hepatitis in Izmir, Turkey. DESIGN AND METHODS: Serum samples from 32 chronic hepatitis patients (21 males, 11 females; mean age 44.2 years, range 23-70 years) with anti-delta positivity were analyzed for hepatitis B and C serologies. After reverse transcription, cDNA of partial delta antigen was amplified by in-house nested PCR. The products of the HDV PCR were bidirectionally sequenced with internal primers using Big Dye Terminator DNA Sequencing Kit (Applied Biosystems, CA, USA) and ABI Prism 310 Genetic Analyzer (Perkin Elmer, USA). Nucleotide sequences of HDV were compared with previously reported sequences and aligned by using ClustalW (1.82). RESULTS: HDV-RNA was positive in 26 (81.3%) of 32 anti-delta positive samples. Comparison of the HDV sequences with published sequences of HDV genotypes I, II, and III indicated that all were closely related to HDV genotype I isolates. Similarity among isolated sequences ranged from 84% to 96%. CONCLUSION: HDV genotyping was successfully performed by direct sequencing of the amplicons obtained from routine HDV-RNA screening PCR tests. All of the HDV isolates from the chronic delta hepatitis patients included in this study were found to be genotype I.

[Bacterial contamination in blood and blood products]. / Kan ve kan urünlerinde bakteriyel kontaminasyon.

Skin disinfection during phlebotomy is a critical step for bacterial contamination of blood and blood products. The aim of this study was to investigate the bacterial contamination rates during phlebotomy and to detect the probable microorganisms present. Skin disinfections of 100 blood donors were performed by using povidone iodine solution with standard procedure. Fifteen mililiters of blood samples were drawn from the transfusion set and inoculated into culture flasks of automated Bact/Alert (BioMerieux) system. Blood cultures were monitorized for one week, and bacteria in positive cultures were identified by using classical microbiological methods in addition with API identification system (BioMerieux; ID32 Staph, 20 Strep). As a result, bacterial growth was detected in four (4%) of the blood samples, whereas 96% of the samples were found sterile. Staphylococcus epidermidis was the microorganism which had been grown in three of the samples, and Streptococcus mutans in one. The positivity rate detected in our study was considered high, since expected bacterial contamination rates in blood transfusions were between 0.2-0.5%. This data indicated that the procedures used in phlebotomy such as the choice of phlebotomy region, disinfectant use and disinfection time should be re-evaluated in our blood centre.

Seroprevalence of hepatitis A,B, and C viruses in Turkish alcoholic cirrhotics and the impact of hepatitis B on clinical profile.

INTRODUCTION: The aims of this study were to detect the seroprevalence of hepatitis A, B, and C viruses in Turkish alcoholic cirrhotics, and to evaluate the impact of hepatitis B infection on clinical profile at first admittance. METHODOLOGY: Serological markers for hepatitis A, B, and C viruses in 300 alcoholic cirrhotics diagnosed between January 1994 and December 2012 were retrospectively reviewed. Among them, 148 eligible patients were divided into group 1 (HBsAg positive, n = 43) and group 2 (HBsAg and anti-HBc negative, n = 105). Clinical characteristics at first admittance of groups 1 and 2 were compared. RESULTS: The seroprevalence of anti-HAV total, HBsAg, and anti-HCV was found to be 91.5%, 16.3%, and 8.2%, respectively. The prevalence of hepatocellular carcinoma was higher in the HbsAg-positive group compared to HbsAg- and anti-HBc-negative group (16.3% vs. 2.9%, p = 0.007). Other clinical features were similar in the two groups. CONCLUSIONS: Alcoholic cirrhotics have higher frequencies of HBsAg and anti-HCV than the general population. These patients should be investigated for coexistent HBV and HCV infections, and HBV vaccination should not be neglected. Alcoholic cirrhotic patients with concomitant HBV infection should be closely screened for hepatocellular carcinoma.

Effect of long-term lamivudine in chronic hepatitis B virus-infected children.

OBJECTIVE: To evaluate, retrospectively, biochemical, serological and histological responses in chronic hepatitis B (CHB)-infected children who received combination therapy and continued with prolonged treatment with lamivudine (3TC). PATIENTS AND METHODS: CHB infection was defined as the presence of hepatitis B surface antigen (HBsAg), hepatitis Be antigen (HBeAg) and hepatitis B virus (HBV) DNA in serum screened at 3-month intervals for at least 1 year, serum alanine aminotransferase (ALT) levels >1.5 times the normal limit and CHB with histological activity index (HAI) >5 by liver biopsy. A total of 99 children with CHB infection were treated with IFN-alpha (three times a week, 5 MU/m2) and 3TC (4 mg/kg/d) orally for 6 months. End of therapy response (CR) was defined as ALT normalization, HBV-DNA clearance and e seroconversion. Partial responders (PR) were defined as patients who had ALT normalization and HBV-DNA clearance, but who had not had e seroconversion. Forty-five children with PR at the end of the sixth month continued to receive 3TC alone thereafter. Breakthrough infection was determined as re-emergence of HBV DNA in serum after its clearance. The response rate, side effects and the breakthrough infection rate were examined on prolonged 3TC treatment. Liver biopsy was held in 29 patients at median 32 (14-66) months of 3TC; pre- and post-treatment liver histology was compared. RESULTS: Pre- and post-treatment evaluation was carried out in 45 children [mean age: 11+/-4.2 years, 31 males (69%), 14 females (31%)] with PR at the end of the sixth month of combination therapy. The initial mean ALT values and HAI scores were 75.6+/-60 IU/l and 8+/-3.3, respectively. 3TC was continued for median 33 (14-66) months and CR was achieved in 15.6% (7/45) and 5.6% (2/36) at the end of first and second year, and 0% (none) at the end of third and fourth year, respectively. Breakthrough incidence was detected in six (13.3%) cases at 12 months and increased to 69.4% (n=25) and 82.4% (n=14) at the end of the second and third years, respectively. Patients with breakthrough continued to receive 3TC. Seroconversion and CR of the mutant virus was achieved in one patient (2.9%) at month 46 of treatment with 3TC. Liver biopsy was held in 29 cases at median 32 (14-66) months of 3TC. Pre- and post-treatment mean HAI scores were 8+/-3.3 and 3.9+/-2.1, respectively (P=0.000). Mean necrosing scores were not different at the beginning and end of therapy (P=1.0). Inflammation, bridging and fibrosis scores decreased to 0.8+/-0.6, 1.3+/-1.2 and 0.6+/-0.8, respectively (P=0.000, P=0.002, P=0.000). CONCLUSION: The long-term 3TC usage in children with PR does not induce complete response and is associated with high breakthrough incidence. However, histological improvement is achieved and/or sustained even in children with HBV DNA breakthrough.