title: Detection of hepatitis B and C viruses in almost all hepatocytes by modified PCR-based in situ hybridization.
abstract: Although PCR-based in situ hybridization (PCR-ISH) can be used to determine the distribution and localization of pathogens in tissues, this approach is hampered by its low specificity. Therefore, we used a highly specific and sensitive PCR-ISH method to reveal the lobular distribution and intracellular localization of hepatitis B virus (HBV) and HCV in chronic liver disease and to clarify the state of persistent HBV and HCV infection in the liver. HBV genomic DNA was detected in almost all hepatocytes, whereas HBV RNA or protein was differentially distributed only in a subset of the HBV DNA-positive region. Further, HCV genomic RNA was detected in almost all hepatocytes and was localized to the cytoplasm. HCV RNA was also detected in the epithelium of the large bile duct but not in endothelial cells, portal tracts, or sinusoidal lymphocytes. In patients with HBV and HCV coinfection, HCV RNA was localized to the noncancerous tissue, whereas HBV DNA was found only in the cancerous tissue. Using this novel PCR-ISH method, we could visualize the staining pattern of HBV and HCV in liver sections, and we obtained results consistent with those of real-time detection (RTD)-PCR analysis. In conclusion, almost all hepatocytes are infected with HBV or HCV in chronic liver disease; this finding implies that the viruses spreads throughout the liver in the chronic stage.
authors.affiliation: Department of Microbiology and Cell Biology, The Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya-ku, Tokyo 156-8506, Japan.
authors.forename: Hideko
authors.initials: H
authors.lastname: Nuriya
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Inoue
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tanaka
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Tsunekazu
authors.initials: T
authors.lastname: Hishima
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Funata
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Kaji
authors.forename: Seishu
authors.initials: S
authors.lastname: Hayashi
authors.forename: Shuichi
authors.initials: S
authors.lastname: Kaneko
authors.forename: Michinori
authors.initials: M
authors.lastname: Kohara
mesh_terms: D000328:Adult; D000368:Aged; D001652:Bile Ducts / Q000821:virology; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D019694:Hepatitis B, Chronic / Q000473:pathology / Q000821:virology*; D019698:Hepatitis C, Chronic / Q000473:pathology / Q000821:virology*; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008099:Liver / Q000821:virology*; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*