title: Lack of evidence for reovirus infection in tissues from patients with biliary atresia and congenital dilatation of the bile duct.
abstract: OBJECTIVE
  To clarify the association between the reovirus infection of the hepatobiliary tree and the development of infantile obstructive cholangiopathy (IOC) including biliary atresia (BA) and congenital dilatation of the bile duct (CBD).
  
  
  METHODS
  We designed reovirus common primers for nested RT-PCR based on the L3 gene segment. The spectrum and the sensitivity of common primers were evaluated with purified reoviral RNAs and reovirus mixed with stool samples. Then, nested RT-PCRs were performed with hepatobiliary and fecal samples obtained from patients with BA, CBD, and control diseases. Additionally, electron microscopy of stool samples was performed.
  
  
  RESULTS
  The L3 common primers could amplify cDNAs synthesized from RNAs of three prototypes of reovirus, and detect as much as 5.0x10(3) plaque forming unit of serotype 3 Dearing strain in 100 mg of fecal samples. However, no amplification product was detected in 136 hepatobiliary tissues taken from 67 patients including 26 BAs and 28 CBDs, or in 65 fecal samples obtained from 15 patients including 10 BAs and 1 CBD. Additionally, viral particles were not found in any stool specimens by the electron microscope.
  
  
  CONCLUSIONS
  These data do not suggest that reoviruses play a major role in the etiology of IOC or BA.
authors.affiliation: Department of Pediatric Surgery (E6), Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-Ku, Chiba, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Saito
authors.forename: Kuniko
authors.initials: K
authors.lastname: Shinozaki
authors.forename: Tadashi
authors.initials: T
authors.lastname: Matsunaga
authors.forename: Tomoko
authors.initials: T
authors.lastname: Ogawa
authors.forename: Takao
authors.initials: T
authors.lastname: Etoh
authors.forename: Toshinori
authors.initials: T
authors.lastname: Muramatsu
authors.forename: Kenji
authors.initials: K
authors.lastname: Kawamura
authors.forename: Hideo
authors.initials: H
authors.lastname: Yoshida
authors.forename: Naomi
authors.initials: N
authors.lastname: Ohnuma
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Shirasawa
mesh_terms: D001652:Bile Ducts / Q000002:abnormalities* / Q000648:ultrastructure; D001656:Biliary Atresia / Q000821:virology*; D005243:Feces / Q000821:virology; D006801:Humans; D007231:Infant, Newborn; D008854:Microscopy, Electron; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012087:Reoviridae / Q000235:genetics / Q000302:isolation & purification*; D012088:Reoviridae Infections / Q000175:diagnosis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral