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Comparison of pulsed-field gel electrophoresis & repetitive sequence-based PCR methods for molecular epidemiological studies of Escherichia coli clinical isolates.

 Il Kwon Bae  ;  Juwon Kim  ;  Je Young Hannah Sun  ;  Seok Hoon Jeong  ;  Yong Rok Kim  ;  Kang Kyun Wang  ;  Kyungwon Lee 
 INDIAN JOURNAL OF MEDICAL RESEARCH, Vol.140(5) : 679-685, 2014 
Journal Title
Issue Date
Bacterial Proteins/genetics ; Bacterial Proteins/isolation & purification* ; Electrophoresis, Gel, Pulsed-Field* ; Escherichia coli/enzymology* ; Escherichia coli/pathogenicity ; Humans ; Molecular Epidemiology ; Polymerase Chain Reaction/methods ; beta-Lactamases/genetics ; beta-Lactamases/isolation & purification*
Escherichia coli ; genetic relationship ; MLST ; PFGE ; rep-PCR ; sequence type
BACKGROUND & OBJECTIVES: PFGE, rep-PCR, and MLST are widely used to identify related bacterial isolates and determine epidemiologic associations during outbreaks. This study was performed to compare the ability of repetitive sequence-based PCR (rep-PCR) and pulsed-field gel electrophoresis (PFGE) to determine the genetic relationships among Escherichia coli isolates assigned to various sequence types (STs) by two multilocus sequence typing (MLST) schemes. METHODS: A total of 41 extended-spectrum β-lactamase- (ESBL-) and/or AmpC β-lactamase-producing E. coli clinical isolates were included in this study. MLST experiments were performed following the Achtman's MLST scheme and the Whittam's MLST scheme, respectively. Rep-PCR experiments were performed using the DiversiLab system. PFGE experiments were also performed. RESULTS: A comparison of the two MLST methods demonstrated that these two schemes yielded compatible results. PFGE correctly segregated E. coli isolates belonging to different STs as different types, but did not group E. coli isolates belonging to the same ST in the same group. Rep-PCR accurately grouped E. coli isolates belonging to the same ST together, but this method demonstrated limited ability to discriminate between E. coli isolates belonging to different STs. INTERPRETATION & CONCLUSIONS: These results suggest that PFGE would be more effective when investigating outbreaks in a limited space, such as a specialty hospital or an intensive care unit, whereas rep-PCR should be used for nationwide or worldwide epidemiology studies.
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1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
Yonsei Authors
Lee, Kyungwon(이경원) ORCID logo https://orcid.org/0000-0003-3788-2134
Jeong, Seok Hoon(정석훈) ORCID logo https://orcid.org/0000-0001-9290-897X
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