Abstract
Between February 2001 and January 2002, an increase in the number of Klebsiella pneumoniae isolates with reduced susceptibility to expanded-spectrum cephalosporins (RSKp) was detected in the neonatal unit of the Juan Canalejo Hospital, and 21 patients were either colonized or infected by the bacterial isolates. The current "gold standard" method for typing K. pneumoniae isolates is pulsed-field gel electrophoresis. However, this technique is expensive and time-consuming. In a search for faster and accurate alternatives to this method, we investigated PCR-based fingerprinting techniques (enterobacterial repetitive intergenic consensus sequence PCR [ERIC-PCR], repetitive extragenic palindromic sequence-based PCR [REP-PCR], and RAPD [randomly amplified polymorphic DNA]) for their ability to characterize K. pneumoniae isolates. The causal agent of the nosocomial outbreak was characterized by these techniques and was found to be a single epidemic strain (RSKp). A multiple regression logistic model was developed to identify potential independent factors associated with colonization and/or infection by RSKp. Logistic regression analysis was applied to all significant variables (P < 0.05) in the univariate analysis, and it was revealed that intubation (odds ratio [OR], 27.0; 95% confidence interval [95%CI], 5.39 to 135.14) and prematurity (OR, 4.4; 95%CI, 0.89 to 21.89) were such independent factors. Moreover, oxime cephalosporins did not appear to be statistically significant. Overall, the results showed that PCR-based techniques are expeditious and useful methods for typing K. pneumoniae isolates. Of the techniques studied, ERIC-PCR showed the highest discriminatory index (D = 0.828), followed by RAPD (D = 0.826) and REP-PCR (D = 0.773)
MeSH Terms
Cephalosporins/pharmacology
Cross Infection/epidemiology,microbiology,prevention & control,transmission
Disease Outbreaks
Electrophoresis, Gel, Pulsed-Field
Feces/microbiology
Humans
Klebsiella Infections/epidemiology,prevention & control,transmission
Klebsiella pneumoniae/drug effects,genetics,growth & development,isolation & purification
Microbial Sensitivity Tests
Polymerase Chain Reaction
Reproducibility of Results
Risk Factors
Sensitivity and Specificity
Spain/epidemiology
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Cartelle Monica
Servicio de Microbiología, Complejo Hospitalario Juan Canalejo, C/As Xubias 84, 15006 La Coruña, Spain.
del Mar Tomas Maria
Pertega Sonia
Beceiro Alejandro
Dominguez M Angeles
Velasco David
Molina Francisca
Villanueva Rosa
Bou German
References (28)
28 references, click to expand
-
Comparison of pulsed-field gel electrophoresis and randomly amplified DNA polymorphism analysis for typing extended-spectrum-beta-lactamase-producing Klebsiella pneumoniae.
J Clin Microbiol. 1996 Oct;34(10):2448-53
PMID: 8880498
-
Effect of an efflux pump inhibitor on the MIC of nalidixic acid for Acinetobacter baumannii and Stenotrophomonas maltophilia clinical isolates.
J Antimicrob Chemother. 2002 Apr;49(4):697-8
PMID: 11909848
-
Molecular epidemiology of infections due to Enterobacter aerogenes: identification of hospital outbreak-associated strains by molecular techniques.
Clin Infect Dis. 1995 Jan;20(1):84-94
PMID: 7727676
-
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 Clin Microbiol. 1998 Sep;36(9):2522-9
PMID: 9705386
-
Validation of use of whole-cell repetitive extragenic palindromic sequence-based PCR (REP-PCR) for typing strains belonging to the Acinetobacter calcoaceticus-Acinetobacter baumannii complex and application of the method to the investigation of a hospital outbreak.
J Clin Microbiol. 1996 May;34(5):1193-202
PMID: 8727902
-
Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing.
J Clin Microbiol. 1995 Sep;33(9):2233-9
PMID: 7494007
-
Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity.
J Clin Microbiol. 1988 Nov;26(11):2465-6
PMID: 3069867
-
A simultaneous outbreak on a neonatal unit of two strains of multiply antibiotic resistant Klebsiella pneumoniae controllable only by ward closure.
J Hosp Infect. 2001 Nov;49(3):183-92
PMID: 11716635
-
Comparison of PCR-ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis for typing Clostridium difficile.
J Clin Microbiol. 2000 Jul;38(7):2484-7
PMID: 10878030
-
Molecular epidemiology of plasmid spread among extended broad-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates in a pediatric hospital.
J Clin Microbiol. 1993 Feb;31(2):179-84
PMID: 8432800
-
Investigation of an apparent cluster of Klebsiella pneumoniae bacteremias using random amplified polymorphic DNA analysis.
Infect Control Hosp Epidemiol. 1996 Nov;17(11):743-5
PMID: 8934243
-
A comparative study of different PCR-based DNA fingerprinting techniques for typing of the Acinetobacter calcoaceticus-A. baumannii complex.
J Med Microbiol. 1996 Jun;44(6):482-9
PMID: 8636966
-
Principles and applications of methods for DNA-based typing of microbial organisms.
J Clin Microbiol. 1999 Jun;37(6):1661-9
PMID: 10325304
-
Outbreak of infection with extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a Mexican hospital.
J Clin Microbiol. 2001 Sep;39(9):3193-6
PMID: 11526149
-
Random amplified polymorphic DNA and plasmid analyses used in investigation of an outbreak of multiresistant Klebsiella pneumoniae.
J Clin Microbiol. 1995 Mar;33(3):713-7
PMID: 7751382
-
Molecular epidemiology of Enterobacter aerogenes acquisition: one-year prospective study in two intensive care units.
J Clin Microbiol. 1996 Jun;34(6):1474-80
PMID: 8735101
-
Transferable plasmid mediating resistance to multiple antimicrobial agents in Klebsiella pneumoniae isolates in Greece.
Clin Microbiol Infect. 2002 Sep;8(9):579-88
PMID: 12427218
-
Diversity of beta-lactam resistance levels among related Klebsiella pneumoniae strains isolated in an intensive care unit.
J Chemother. 2000 Jun;12(3):204-7
PMID: 10877514
-
AcrAB Efflux System: Expression and Contribution to Fluoroquinolone Resistance in Klebsiella spp.
Antimicrob Agents Chemother. 2002 Dec;46(12):3984-6
PMID: 12435706
-
Characterization of Shiga toxin-producing Escherichia coli O157:H7 isolated in Italy and in France.
J Clin Microbiol. 2002 Dec;40(12):4619-24
PMID: 12454162
-
Outbreak of a multiresistant Klebsiella pneumoniae strain in an intensive care unit: antibiotic use as risk factor for colonization and infection.
Clin Infect Dis. 2000 Jan;30(1):55-60
PMID: 10619733
-
Distribution of extended-spectrum beta-lactamases in clinical isolates of Enterobacteriaceae in Vietnam.
Antimicrob Agents Chemother. 2002 Dec;46(12):3739-43
PMID: 12435670
-
Epidemiological study by pulsed-field gel electrophoresis of an outbreak of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a geriatric hospital.
J Clin Microbiol. 1994 Feb;32(2):301-5
PMID: 8150938
-
Characteristics of bacteremia between community-acquired and nosocomial Klebsiella pneumoniae infection: risk factor for mortality and the impact of capsular serotypes as a herald for community-acquired infection.
Arch Intern Med. 2002 May 13;162(9):1021-7
PMID: 11996612
-
RAPD typing of Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens and Pseudomonas aeruginosa isolates using standardized reagents.
Clin Microbiol Infect. 1999 May;5(5):270-276
PMID: 11856266
-
Identification of a Klebsiella pneumoniae strain associated with nosocomial urinary tract infection.
J Clin Microbiol. 1997 Sep;35(9):2370-4
PMID: 9276418
-
Duodenal microflora in very-low-birth-weight neonates and relation to necrotizing enterocolitis.
J Clin Microbiol. 2000 Dec;38(12):4539-47
PMID: 11101593
-
Study of an outbreak of cefoxitin-resistant Klebsiella pneumoniae in a general hospital.
J Clin Microbiol. 1997 Feb;35(2):508-10
PMID: 9003629