title: Evaluation of small-subunit rRNA touchdown polymerase chain reaction for direct detection of Entamoeba histolytica in human pus samples from patients with amoebic liver abscess.
abstract: OBJECTIVE
  The aim of the present study was to evaluate the use of touchdown polymerase chain reaction (TD-PCR) for the detection of Entamoeba histolytica in liver pus samples obtained from patients with a clinical diagnosis of amoebic liver abscess (ALA) using small-subunit rRNA (SSU rRNA) as the target gene.
  
  
  METHODS
  Microscopic examination in vitro culture and serological test for the detection of E. histolytica in 67 pus samples obtained from ALA patients was performed. Molecular studies were carried out by both conventional PCR and TD-PCR targeting the SSU rRNA gene using the same sets of primers and the results were compared.
  
  
  RESULTS
  TD-PCR detected the presence of E. histolytica in 86.5% of the liver pus samples within 2.5 h as compared with 82.08% by conventional PCR within 3.5-4 h.
  
  
  CONCLUSIONS
  TD-PCR assay may serve as a relatively better detection method for E. histolytica over conventional PCR with respect to the turnaround time, increased sensitivity, specificity and yield.
authors.affiliation: Department of Microbiology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110 029, India.
authors.forename: P
authors.initials: P
authors.lastname: Singh
authors.forename: B R
authors.initials: BR
authors.lastname: Mirdha
authors.forename: V
authors.initials: V
authors.lastname: Ahuja
authors.forename: S
authors.initials: S
authors.lastname: Singh
mesh_terms: D019411:Clinical Laboratory Techniques / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004748:Entamoeba histolytica / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008101:Liver Abscess, Amebic / Q000175:diagnosis* / Q000469:parasitology; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016053:RNA, Protozoan / Q000235:genetics*; D012337:RNA, Ribosomal, 18S / Q000235:genetics*; D012680:Sensitivity and Specificity; D013492:Suppuration / Q000469:parasitology*; D013997:Time Factors
chemical_list: D017931:DNA Primers; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S