title: Using the polymerase chain reaction coupled with denaturing gradient gel electrophoresis to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted acute severe pancreatitis.
abstract: OBJECTIVE
  To investigate the use of PCR and DGGE to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted severe AP.
  
  
  METHODS
  Patients with biochemical and clinical evidence of acute pancreatitis and an APACHE II score > or = 8 were enrolled. PCR and DGGE were employed to detect bacterial translocation in blood samples collected on d 1, 3, and 8 after the admission. Standard microbial blood cultures were taken when there was clinical evidence of sepsis or when felt to be clinically indicated by the supervising team.
  
  
  RESULTS
  Six patients were included. Of all the patients investigated, only one developed septic complications; the others had uneventful illness. Bacteria were detected using PCR in 4 of the 17 collected blood samples. The patient with sepsis was PCR-positive in two samples (taken on d 1 and 3), despite three negative blood cultures. Using DGGE and specific primers, the bacteria in all blood specimens which tested positive for the presence of bacterial DNA were identified as E coli.
  
  
  CONCLUSIONS
  Our study confirmed that unlike traditional microbiological techniques, PCR can detect the presence of bacteria in the blood of patients with severe AP. Therefore, this latter method in conjunction with DGGE is potentially an extremely useful tool in predicting septic morbidity and evaluating patients with the disease. Further research using increased numbers of patients, in particular those patients with necrosis and sepsis, is required to assess the reliability of PCR and DGGE in the rapid diagnosis of infection in AP.
authors.affiliation: Gastroenterology, Diagnostic Procedures Unit, Fremantle Hospital, Fremantle, Western Australia, WA 6011, Australia. pearcey@screaming.net
authors.forename: Callum B
authors.initials: CB
authors.lastname: Pearce
authors.forename: Vitaly
authors.initials: V
authors.lastname: Zinkevich
authors.forename: Iwona
authors.initials: I
authors.lastname: Beech
authors.forename: Viera
authors.initials: V
authors.lastname: Funjika
authors.forename: Ana Garcia
authors.initials: AG
authors.lastname: Ruiz
authors.forename: Afraa
authors.initials: A
authors.lastname: Aladawi
authors.forename: Hamish D
authors.initials: HD
authors.lastname: Duncan
mesh_terms: D000208:Acute Disease; D001419:Bacteria / Q000302:isolation & purification; D001483:Base Sequence; D004269:DNA, Bacterial / Q000097:blood* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D006801:Humans; D010195:Pancreatitis / Q000150:complications* / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D018746:Systemic Inflammatory Response Syndrome / Q000150:complications* / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial