학술논문
Identification of AcinetobacterSpecies and Genotyping of Acinetobacter baumanniiby Multilocus PCR and Mass Spectrometry
Document Type
Article
Author
Ecker, Joseph A.; Massire, Christian; Hall, Thomas A.; Ranken, Raymond; Pennella, Thuy-Trang D.; Agasino Ivy, Cristina; Blyn, Lawrence B.; Hofstadler, Steven A.; Endy, Timothy P.; Scott, Paul T.; Lindler, Luther; Hamilton, Tacita; Gaddy, Charla; Snow, Kerry; Pe, Marie; Fishbain, Joel; Craft, David; Deye, Gregory; Riddell, Scott; Milstrey, Eric; Petruccelli, Bruno; Brisse, Sylvain; Harpin, Vanessa; Schink, Amy; Ecker, David J.; Sampath, Rangarajan; Eshoo, Mark W.
Source
Journal of Clinical Microbiology; August 2006, Vol. 44 Issue: 8 p2921-2932, 12p
Subject
Language
ISSN
00951137; 1098660X
Abstract
ABSTRACTMembers of the genus Acinetobacterare ubiquitous in soil and water and are an important cause of nosocomial infections. A rapid method is needed to genotype Acinetobacterisolates to determine epidemiology and clonality during infectious outbreaks. Multilocus PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) is a method that uses the amplicon base compositions to genotype bacterial species. In order to identify regions of the Acinetobactergenome useful for this method, we sequenced regions of six housekeeping genes (trpE, adk, efp, mutY, fumC, and ppa) from 267 isolates of Acinetobacter. Isolates were collected from infected and colonized soldiers and civilians involved in an outbreak in the military health care system associated with the conflict in Iraq, from previously characterized outbreaks in European hospitals, and from culture collections. Most of the isolates from the Iraqi conflict were Acinetobacter baumannii(189 of 216 isolates). Among these, 111 isolates had genotypes identical or very similar to those associated with well-characterized A. baumanniiisolates from European hospitals. Twenty-seven isolates from the conflict were found to have genotypes representing different Acinetobacterspecies, including 8 representatives of Acinetobactergenomospecies 13TU and 13 representatives of Acinetobactergenomospecies 3. Analysis by the PCR/ESI-MS method using nine primer pairs targeting the most information-rich regions of the trpE, adk, mutY, fumC, and ppagenes distinguished 47 of the 48 A. baumanniigenotypes identified by sequencing and identified at the species level at least 18 Acinetobacterspecies. Results obtained with our genotyping method were essentially in agreement with those obtained by pulse-field gel electrophoresis analysis. The PCR/ESI-MS genotyping method required 4 h of analysis time to first answer with additional samples subsequently analyzed every 10 min. This rapid analysis allows tracking of transmission for the implementation of appropriate infection control measures on a time scale previously not achievable.