Standardization and Interlaboratory Reproducibility Assessment of Pulsed-Field Gel Electrophoresis-Generated Fingerprints of Acinetobacter baumannii

• DOI: 10.1128/jcm.43.9.4328-4335.2005
• Publication Date: 2005-09-06T21:58:02Z
• Authors: Harald Seifert; Lucilla Dolzani; Raffaela Bressan; Tanny van der Rei...; Beppie van Strijen; Danuta Stefanik; Herre Heersma; Lenie Dijkshoorn
• Keywords: Acinetobacter baumannii; DNA profiling

ABSTRACT A standard procedure for pulsed-field gel electrophoresis (PFGE) of macrorestriction fragments Acinetobacter baumannii was set up and validated its interlaboratory reproducibility potential use in the construction an Internet-based database international monitoring epidemic strains. The PFGE fingerprints strains were generated at three different laboratories with ApaI as restriction enzyme by a rigorously standardized procedure. results analyzed respective also centrally national reference institute. In first phase study, 20 A. strains, including 3 isolates each from well-characterized hospital outbreaks 11 sporadic distributed blindly to participating laboratories. local groupings laboratory identical allowed identification epidemiologically related belonging clusters identified all unrelated distinct. Central pattern analysis using band-based Dice coefficient unweighted pair group method mathematical averaging clustering algorithm showed 95% matching outbreak processed 87% corresponding if they second 30 representing 10 parts Europe (3 per outbreak) laboratories, so that investigated independent isolates. computer-assisted cluster correctly according their threshold. conclusion, enabled us generate locations sufficient electronic monitor geographic spread