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Evaluation of Three Molecular Typing Techniques for Nonfermentative Gram-Negative Bacilli

Published online by Cambridge University Press:  02 January 2015

Suzane Silbert ,
Michael A. Pfaller ,
Richard J. Hollis ,
Afonso L. Barth  and
Hélio S. Sader
Suzane Silbert*
Affiliation:
Laboratório Especial de Microbiologia Clínica, Disciplina de Doenças Infecciosas e Parasitàrias, Universidade Federal de São Paulo, São Paulo, Brasil
Michael A. Pfaller
Affiliation:
Molecular Epidemiology and Fungus Testing Laboratory, Department of Pathology, University of Iowa College of MedicineIowa City, Iowa
Richard J. Hollis
Affiliation:
Molecular Epidemiology and Fungus Testing Laboratory, Department of Pathology, University of Iowa College of MedicineIowa City, Iowa
Afonso L. Barth
Affiliation:
Unidade de Pesquisa Biomédica, Serviço de Patologia Clínica, Hospital de Clínicas de Porto Alegre, Porto Alegre, Brasil
Hélio S. Sader
Affiliation:
Laboratório Especial de Microbiologia Clínica, Disciplina de Doenças Infecciosas e Parasitàrias, Universidade Federal de São Paulo, São Paulo, Brasil
*
Laboratòrio Especial de Microbiologia Clínica (LEMC)Disciplina de Doenças Infecciosas e Parasitárias, Escola Paulista de Medicina – Universidade Federal de São Paulo, Rua Leandro Dupret, 188, São Paulo, SP – CEP 04025-010, Brasil
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Abstract

Objective:

To evaluate three different DNA techniques for typing nonfermentative gram-negative bacilli isolated from Latin American hospitals.

Design:

One hundred twenty-six nonfermentative gram-negative bacilli were typed.

Participants:

Pseudomonas aeruginosa (n = 64) and Acinetobacter baumannii (n = 42) samples were obtained from blood cultures of patients admitted to 10 medical centers in Latin America during 1998 and Stenotrophomonas maltophilia (n = 20) samples were obtained from patients admitted to the Hospital São Paulo between 1999 and 2001.

Methods:

All samples were typed using automated ribotyping, PFGE, and ERIC-PCR. The discriminatory power for each technique was calculated using Hunter's generalized formula.

Results:

All strains could be typed by automated ribotyping and ERIC-PCR, but two strains (1.6%) were not typeable by PFGE. All three techniques showed 100% reproducibility. The time to obtain the results was shorter for automated ribotyping and ERIC-PCR compared with PFGE. Likewise, the costs for ERIC-PCR and PFGE were lower than those for automated ribotyping. The interpretation of results was more complicated and more difficult with ERIC-PCR than with both PFGE and automated ribotyping. All techniques presented excellent discriminatory power for P. aeruginosa (0.98). PFGE presented the highest discriminatory power (0.94) for A. baumannii, and both PFGE and ERIC-PCR showed higher discriminatory power (0.90 for both) than automated ribotyping (0.82) for S. maltophilia.

Conclusions:

PFGE showed the highest discriminatory power for typing these nonfermentative gram-negative bacilli. However, automated ribotyping and ERIC-PCR can provide results in a shorter time period with similar discriminatory power.

Type
Orginal Articles
Information
Infection Control & Hospital Epidemiology , Volume 25 , Issue 10 , October 2004 , pp. 847 - 851
Copyright
Copyright © The Society for Healthcare Epidemiology of America 2004 

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References

1. Hancock, RE. Resistance mechanisms in Pseudomonas aeruginosa and other nonfermentative gram-negative bacteria. Clin Infect Dis 1998;27:S93–S99.10.1086/514909Google Scholar
2. Pfaller, MA, Acar, J, Jones, RN, Verhoef, J, Turnidge, J, Sader, HS. Integration of molecular characterization of microorganisms in a global antimicrobial resistance surveillance program. Clin Infect Dis 2001;32:S104–S113.Google Scholar
3. Struelens, MJ, Gheldre, Y, De Deplano, A. Comparative and library epidemiological typing systems; outbreak investigation versus surveillance systems. Infect Control Hosp Epidemiol 1998;19:565–569.10.2307/30141781Google Scholar
4. Pfaller, MA. Molecular epidemiology in the care of patients. Arch Pathol Lab Med 1999;123:1007–1010.Google Scholar
5. Bruce, J. Automated system rapidly identifies and characterizes microorganisms in food. Food Technology 1996;50:77–81.Google Scholar
6. Thompson, CJ, Daly, C, Barrett, TJ, Getchell, JP, Gilchrist, MJ, Loeffelholz, MJ. Insertion element IS3-based PCR method for subtyping Escherichia coli 0157:H7 J Clin Microbiol 1998;36:1180–1184.Google Scholar
7. Olive, MD, Bean, P. Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol 1999;37:1661–1669.Google Scholar
8. Gales, AC, Jones, RN, Turnidge, R, Rennie, R, Ramphai, R. Characterization of Pseudomonas aeruginosa isolates: occurrence rates, antimicrobial susceptibility patterns, and molecular typing in the global SENTRY Antimicrobial Surveillance Program, 1997-1999. Clin Infect Dis 2001a;32:S146–S155.Google Scholar
9. Gales, AC, Jones, RN, Forward, KR, Liftares, J, Sader, HS, Verhoef, J. Emerging importance of multidrug-resistant Acinetobacter species and Stenotrophomonas maltophilia as pathogens in seriously ill patients: geographic patterns, epidemiological features, and trends in the SENTRY Antimicrobial Surveillance Program, 1997-1999. Clin Infect Dis 2001b;32:S104–S113.Google Scholar
10. Pfaller, MA, Hollis, RJ, Sader, HS. Molecular biology: PFGE analysis of chromosomal restriction fragments. In: Isenberg, HD, ed. Clinical Microbiology Procedures Handbook. Washington, DC: ASM Press; 1992:10.5.c.1–10.5.c.11.Google Scholar
11. Versalovic, J, Koeuth, T, Lupski, JR. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res 1991;19:6823–6831.10.1093/nar/19.24.6823Google Scholar
12. Tenover, FC, Arbeit, RD, Goering, RV. 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;18:426–439.10.2307/30141252Google Scholar
13. Viedema, DG, Marin, M, Cercenado, E, Alonso, R, Rodriguez-Créixems, M, Bouza, E. Evidence of nosocomial Stenotrophomonas maltophilia cross-infection in a neonatology unit analyzed by three molecular typing methods. Infect Control and Hosp Epidemiol 1999;20:816–820.Google Scholar
14. Berg, G, Roskot, N, Smalla, K. Genotypie and phenotypic relationships between clinical and environmental isolates of Stenotrophomonas maltophilia . J Clin Microbiol 1999;37:3594–3600.Google Scholar
15. D'Agata, EM, Gerrits, MM, Tang, YW, Samore, M, Kusters, JG. Comparison of pulsed-field gel electrophoresis and amplified fragment-length polymorphism for epidemiological investigations of common nosocomial pathogens. Infect Control Hosp Epidemiol 2001;22:550–554.Google Scholar
16. Pellegrino, FL, Teixeira, LM, Carvalho, MG, et al. Occurrence of a multidrug-resistant Pseudomonas aeruginosa clone in different hospitals in Rio de Janeiro, Brazil. J Clin Microbiol 2002;40:2420–2424.Google Scholar
17. Hunter, PR. Reproducibility and indices of discriminatory power of microbial typing methods. J Clin Microbiol 1990;28:1902–1905.Google Scholar
18. Hollis, RJ, Bruce, JL, Fritschel, JS, Pfaller, MA. Comparative evaluation of an automated ribotyping instrument versus pulsed-field gel electrophoresis for epidemiological investigation of clinical isolates of bacteria. Diagn Microbiol Infect Dis 1999;34:263–268.10.1016/S0732-8893(99)00033-4Google Scholar
19. Dambaugh, TR, Mangiaterra, E, Fritschel, SJ. Ribotype characterization of Salmonella and E. coli 0157:H7 with Prall on the Riboprinter Microbial Characterization System. In: Abstracts of the General Meeting of the American Society for Microbiology. Miami, FL: American Society for Microbiology; 1997. Abstract P55:14.Google Scholar
20. Grundmann, H, Schneider, C, Hartung, D, Daschner, FD, Pitt, TL. Discriminatory power of three DNA-based typing techniques for Pseudomonas aeruginosa . J Clin Microbiol 1995;33:528–534.Google Scholar
21. Romling, U, Tummler, B. Achieving 100% typeability of Pseudomonas aeruginosa by pulsed-field gel electrophoresis. J Clin Microbiol 2000;38:464–465.10.1128/JCM.38.1.464-465.2000Google Scholar
22. Silbert, S, Boyken, L, Hollis, RJ, Pfaller, MA. Improving typeability by pulsed-field gel electrophoresis. Diagn Microbiol Infect Dis 2003;47:619–621.10.1016/S0732-8893(03)00164-0Google Scholar