Hostname: page-component-586b7cd67f-g8jcs Total loading time: 0 Render date: 2024-11-28T16:02:56.535Z Has data issue: false hasContentIssue false

Comparison of Pulsed-Field Gel Electrophoresis and Amplified Fragment-Length Polymorphism for Epidemiological Investigations of Common Nosocomial Pathogens

Published online by Cambridge University Press:  02 January 2015

Erika M.C. D'Agata*
Affiliation:
Division of Infectious Diseases, Vanderbilt University, Nashville, Tennessee
Monique M. Gerrits
Affiliation:
Department of Medical Microbiology, Free University, Amsterdam, The Netherlands
Yi-Wei Tang
Affiliation:
Division of Infectious Diseases, Vanderbilt University, Nashville, Tennessee
M. Samore
Affiliation:
Division of Infectious Diseases, IDS Hospital, Salt Lake City, Utah
Johannes G. Kusters
Affiliation:
Department of Medical Microbiology, Free University, Amsterdam, The Netherlands
*
Beth Israel Deaconess Medical Center, Division of Infectious Diseases, Kennedy 6A, 330 Brookline Ave, Boston, MA 02215

Abstract

Objective:

To compare molecular typing by amplified fragment-length polymorphism (AFLP) analysis with pulsed-field gel electrophoresis (PFGE) with respect to the ability to differentiate between epidemiologically related and unrelated isolates of common nosocomial pathogens recovered during a period of endemicity.

Design:

Retrospective laboratory analysis.

Setting:

Tertiary-care institution.

Methods:

17 isolates of Acinetobacter baumannii, 22 isolates of Pseudomonas aeruginosa, and 22 vancomycin-resistant Enterococcus faecium (VRE) were typed by both methods.

Results:

AFLP generated comparable results to PFGE for A baumannii and P aeruginosa isolates; both methods identified epidemiologically related and unrelated isolates. However, strain typing of VRE isolates produced discordant results between the two methods. PFGE identified 10 different strain types and differentiated between all epidemiologically related and unrelated isolates. In contrast, AFLP generated only five different strain types, three of which contained both epidemiologically related and unrelated isolates.

Conclusion:

Molecular typing by AFLP is comparable to PFGE for A baumannii and P aeruginosa isolates. For VRE isolates, however, PFGE remains the method of choice.

Type
Original Articles
Copyright
Copyright © The Society for Healthcare Epidemiology of America 2001

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1.Tenover, FC, Arbeit, RD, Goering, RV. The Molecular Typing Working Group of the Society for Healthcare Epidemiology of America. How to select and interpret molecular strain typing methods for epidemiological studies of bacterial infections: a review for healthcare epidemiologists. Infect Control Hosp Epidemiol 1997;18:426439.CrossRefGoogle Scholar
2.Olive, DM, Bean, P. Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol 1999;37:16611669.CrossRefGoogle Scholar
3.Janssen, P, Coopman, R, Huys, G, Swings, J, Bleeker, M, Vos, P, et al. Evaluation of the DNA fingerprinting method AFLP as a new tool in bacterial taxonomy. Microbiology 1996;142:18811893.Google Scholar
4.Vos, P, Hogers, R, Bleeker, M, Reijans, M, Van de Lee, T, Homes, M, et al. AFLP: a new technique for DNA fingerprinting. Nucl Acids Res 1995;23:44074414.CrossRefGoogle ScholarPubMed
5.Dijkshoorn, L, Aucken, H, Gerner-Smidt, P, Janssen, P, Kaufmann, ME, Garaizar, J, et al. Comparison of outbreak and nonoutbreak Acinetotecter baumannii strains by genotypic and phenotypic methods. J Clin Microbiol 1006;34:15191525.CrossRefGoogle Scholar
6.Gibson, JR, Slater, E, Xery, J, Tompkins, DA, Owen, RJ. Use of an amplified-fragment length polymorphism technique to fingerprint and differentiate isolates of Helicobacter pylori. J Clin Microbiol 1998;36:25802585.Google Scholar
7.Grady, R, Desai, M, O'Neill, G, Cookson, B, Stanely, J. Genotyping of epidemic methicillin-resistant Staphylococcus aureus phage type 15 isolates by fluorescent amplified-fragment length polymorphism analysis. J Qin Microbiol 1999;37:31983203.Google ScholarPubMed
8.Huys, G, Coopman, R, Janssen, P, Kersters, K. High-resolution genotypic analysis of the genus Aeromonas by AFLP fingerprinting. Int J Syst Bacterid 1996;46:572580.CrossRefGoogle ScholarPubMed
9.Janssen, P, Dijkshoorn, L. High resolution DNA fingerprinting of Acinetobacter outbreak strains. FEMS Microbiol Lett 1996;142:191194.Google Scholar
10.Kiem, P, Kalif, A, Schupp, J, Hill, K, Travis, SE, Richmond, K, et al. Molecular evolution and diversity in Bacillus antracis as detected by amplified length polymorphism markers. J Bacteriol 1997;179:818824.CrossRefGoogle Scholar
11.Koelman, JGM, Stoof, J, Biesmans, DJ, Savelkoul, PHM, Vandenbroucke-Grauls, CMJ. Comparison of amplified ribosomal DNA restriction analysis, random amplified polymorphic DNA analysis, and amplified fragment length polymorphism fingerprinting for identification of Acinetobacter genomic species and typing of Acinetobacter baumannii. J Qin Microbiol 1998;36:25222529.Google Scholar
12.Rcardeau, M, Prod'Hom, G, Raskine, L, LePennec, MP, Vincent, V. Genotypic characterization of five subspecies of Mycobacterium kansasii. J Clin Microbiol 1997;35:2532.Google Scholar
13.van Eldere, J, Janssen, P, Hoefnagels-Schuermans, A, van Lierde, S, Peetermans, WE. Amplified-fragment length polymorphism analysis versus macro-restriction fragment analysis for molecular typing of Streptococcus pneumoniae isolates. J Qin Microbiol 1999;37:20532057.Google Scholar
14.Speijer, H, Savelkoul, PHM, Bonten, MJ, Stobberingh, EE, Tjhie, JHT. Application of different genotyping methods for Pseudomonas aeruginosa in a setting of endemicity in an intensive care unit. J Clin Microbiol 1999;37:36543661.Google Scholar
15.D'Agata, E, Venkataraman, L, DeGirolami, P, Samore, M. The molecular epidemiology of acquisition of ceftazidime-resistant gram-negative strains in a non-outbreak setting. J Qin Microbiol 1997;35:26022605.Google Scholar
16.D'Agata, EMC, Schaffner, W, Ii, H, Tang, YW. Genomic variability of vancomycin-resistantenterococci: inter- and intra-patient analysis. Qin Infect Dis 1999;29:991.Google Scholar
17.D'Agata, EMC, Venkataraman, L, DeGirolami, P, Samore, M. The molecular epidemiology of ceftazidime-resistant gram-negative bacilli on inanimate surfaces and their role in cross-transmission during non-outbreak periods. J Qin Microbiol 1999;37:30653067.Google Scholar
18.Ostrowski, B, Venkataraman, L, D'Agata, EMC, Gold, H, DeGirolami, P, Samore, M. Vancomycin-resistant Enterococci in intensive care units: high frequency of stool carriage during non-outbreak periods. Arch Intern Med 1999;159:14671472.Google Scholar
19.Tenover, FC, Arbeit, RD, Goering, RV, Mickelsen, PA, Murray, BE, Persing, DH, et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Qin Microbiol 1995;33:22232229.Google Scholar
20.Hunter, PR, Gaston, MA. Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Qin Microbiol 1988;26:24652466.Google Scholar
21.Knight, JC, Udalova, I, Hill, AVS, Greenwood, BM, Peshu, N, March, KD, et al. A polymorphism that affects OCT-1 binding to the TNF promoter region is associated with severe malaria. Nature 1999;22:145150.Google Scholar