Interlaboratory reproducibility of DiversiLab rep-PCR typing and clustering of Acinetobacter baumannii isolates

J Med Microbiol. 2012 Jan;61(Pt 1):137-141. doi: 10.1099/jmm.0.036046-0. Epub 2011 Sep 8.

Abstract

We have investigated the reproducibility of DiversiLab rep-PCR fingerprints between two laboratories with the aim of determining if the fingerprints and clustering are laboratory-specific or portable. One-hundred non-duplicate Acinetobacter baumannii isolates were used in this study. DNA isolation and rep-PCR were each performed separately in two laboratories and rep-PCR patterns generated in laboratory A were compared with those from laboratory B. Twelve A. baumannii isolates processed in laboratory A showed ≥98 % pattern similarity with the corresponding 12 isolates tested in laboratory B and were considered identical. Sixty-four isolates showed 95-97.9 % similarity with their corresponding isolates. Twenty-three isolates showed 90-94 % similarity with the corresponding isolates, while one isolate showed only 87.4 % similarity. However, intra-laboratory clustering was conserved: isolates that clustered in laboratory A also clustered in laboratory B. While clustering was conserved and reproducible at two different laboratories, demonstrating the robustness of rep-PCR, interlaboratory comparison of individual isolate fingerprints showed more variability. This comparison allows conclusions regarding clonality to be reached independent of the laboratory where the analysis is performed.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acinetobacter Infections / epidemiology*
  • Acinetobacter Infections / microbiology
  • Acinetobacter baumannii / classification*
  • Acinetobacter baumannii / genetics
  • Acinetobacter baumannii / isolation & purification
  • Bacterial Typing Techniques*
  • Cluster Analysis*
  • DNA Fingerprinting / methods
  • DNA, Bacterial / analysis
  • DNA, Bacterial / genetics
  • Humans
  • Laboratories
  • Polymerase Chain Reaction / methods*
  • Repetitive Sequences, Nucleic Acid / genetics*
  • Reproducibility of Results

Substances

  • DNA, Bacterial