title: Nested polymerase chain reaction in the diagnosis of negative Ziehl-Neelsen stained Mycobacterium tuberculosis fistula-in-ano: report of four cases.
abstract: OBJECTIVE
  Mycobacterium is one of the causes of granulomatous diseases within the anorectal region. Early diagnosis of infection is important before the use of antituberculosis chemotherapy. Clinical diagnosis is usually dependent on microscopic detection using Ziehl-Neelsen stain and mycobacterial culture, but the sensitivity and specificity of these two methods are low. In this study nested polymerase chain reaction was used to detect mycobacterial infection in anal fistulas.
  
  
  METHODS
  Paraffin-embedded specimens from three patients and discharge from one patient were used. DNA extraction was performed using phenol/chloroform techniques. IS6110-based nested polymerase chain reaction, yielding a 259-bp amplicon, for the diagnosis of infection was done to facilitate treatment.
  
  
  RESULTS
  Four cases of suspected fistulas-in-ano presented with persistent fistula or unhealed wound. Histopathologic examination revealed granulomatous inflammation with failed microscopic detection of acid-fast bacilli using Ziehl-Neelsen stain. Nested polymerase chain reaction confirmed the presence of in all cases. The anal lesions healed rapidly following a course of antituberculosis therapy.
  
  
  CONCLUSIONS
  Molecular diagnosis of fistula-in-ano by nested polymerase chain reaction is useful for clinically highly suspected infection despite a negative Ziehl-Neelsen stain.
authors.affiliation: Department of Surgery, College of Medicine, National Cheng Kung University Hospital, Tainan, Taiwan.
authors.forename: Yan-Shen
authors.initials: YS
authors.lastname: Shan
authors.forename: Jing-Jou
authors.initials: JJ
authors.lastname: Yan
authors.forename: Edgar D
authors.initials: ED
authors.lastname: Sy
authors.forename: Ying-Tai
authors.initials: YT
authors.lastname: Jin
authors.forename: Jenq-Chang
authors.initials: JC
authors.lastname: Lee
mesh_terms: D000368:Aged; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000472:pathogenicity; D016133:Polymerase Chain Reaction / Q000379:methods*; D012003:Rectal Fistula / Q000150:complications / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014385:Tuberculosis, Gastrointestinal / Q000175:diagnosis* / Q000473:pathology
chemical_list: D004269:DNA, Bacterial