title: Distinct distribution pattern of hepatitis B virus genotype C and D in liver tissue and serum of dual genotype infected liver cirrhosis and hepatocellular carcinoma patients.
abstract: OBJECTIVE
  The impact of co-infection of several hepatitis B virus (HBV) genotypes on the clinical outcome remains controversial. This study has for the first time investigated the distribution of HBV genotypes in the serum and in the intrahepatic tissue of liver cirrhotic (LC) and hepatocellular carcinoma (HCC) patients from India. In addition, the genotype-genotype interplay and plausible mechanism of development of HCC has also been explored.
  
  
  METHODS
  The assessment of HBV genotypes was performed by nested PCR using either surface or HBx specific primers from both the circulating virus in the serum and replicative virus that includes covalently closed circular DNA (cccDNA) and relaxed circular DNA (rcDNA) of HBV from the intrahepatic tissue. The integrated virus within the host chromosome was genotyped by Alu-PCR method. Each PCR products were cloned and sequences of five randomly selected clones were subsequently analysed.
  
  
  RESULTS
  HBV/genotype D was detected in the serum of all LC and HCC patients whereas the sequences of the replicative HBV DNA (cccDNA and rcDNA) from the intrahepatic tissue of the same patients revealed the presence of both HBV/genotype C and D. The sequences of the integrated viruses exhibited the solo presence of HBV/genotype C in the majority of LC and HCC tissues while both HBV/genotype C and D clones were found in few patients in which HBV/genotype C was predominated. Moreover, compared to HBV/genotype D, genotype C had higher propensity to generate double strand breaks, ER stress and reactive oxygen species and it had also showed higher cellular homologous-recombination efficiency that engendered more chromosomal rearrangements, which ultimately led to development of HCC.
  
  
  CONCLUSIONS
  Our study highlights the necessity of routine analysis of HBV genotype from the liver tissue of each chronic HBV infected patient in clinical practice to understand the disease prognosis and also to select therapeutic strategy.
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Somenath
authors.initials: S
authors.lastname: Datta
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Shrabasti
authors.initials: S
authors.lastname: Roychoudhury
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Alip
authors.initials: A
authors.lastname: Ghosh
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Debanjali
authors.initials: D
authors.lastname: Dasgupta
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Amit
authors.initials: A
authors.lastname: Ghosh
authors.forename: Bidhan Chandra
authors.initials: BC
authors.lastname: Chakraborty
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Bidhan
authors.initials: B
authors.lastname: Chakraborty
authors.forename: Sukanta
authors.initials: S
authors.lastname: Ray
authors.affiliation: Department of Gastro-Intestinal surgery, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Sukanta
authors.initials: S
authors.lastname: Roy
authors.affiliation: Centre for Liver and Biliary Surgery, Indraprastha Apollo Hospital, New Delhi, India.
authors.forename: Subash
authors.initials: S
authors.lastname: Gupta
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Amal Kumar
authors.initials: AK
authors.lastname: Santra
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Simanti
authors.initials: S
authors.lastname: Datta
authors.affiliation: Department of Hepatology, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Kausik
authors.initials: K
authors.lastname: Das
authors.affiliation: Department Gastroenterology, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Gopal Krishna
authors.initials: GK
authors.lastname: Dhali
authors.affiliation: Department of Hepatology, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Abhijit
authors.initials: A
authors.lastname: Chowdhury
authors.affiliation: Centre for Liver Research, School of Digestive and Liver Diseases, Institute of Post Graduate Medical Education and Research, Kolkata, India.
authors.forename: Soma
authors.initials: S
authors.lastname: Banerjee
mesh_terms: D000328:Adult; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000401:mortality / Q000473:pathology / Q000821:virology*; D045744:Cell Line, Tumor; D060085:Coinfection; D053903:DNA Breaks, Double-Stranded; D004270:DNA, Circular / Q000097:blood / Q000235:genetics; D004279:DNA, Viral / Q000097:blood / Q000235:genetics; D005838:Genotype; D056945:Hep G2 Cells; D006513:Hepatitis B e Antigens / Q000097:blood / Q000276:immunology; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000235:genetics / Q000821:virology*; D006801:Humans; D007194:India; D008099:Liver / Q000473:pathology / Q000821:virology; D008103:Liver Cirrhosis / Q000401:mortality / Q000473:pathology / Q000821:virology*; D008113:Liver Neoplasms / Q000401:mortality / Q000473:pathology / Q000821:virology*; D016031:Liver Transplantation; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D011379:Prognosis; D017382:Reactive Oxygen Species / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D004270:DNA, Circular; D004279:DNA, Viral; D006513:Hepatitis B e Antigens; D017382:Reactive Oxygen Species