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Comparison of PCR-RFLP and PFGE for determining the clonality of enterohemorrhagic Escherichia coli strains

Abstract We report here on a comparative evaluation of PCR-restriction fragment length polymorphism (PCR-RFLP) and pulsed-field gel electrophoresis (PFGE) assays, and ascertain the clonal relationship between 13 enterohemorrhagic Escherichia coli O157 : H7 strains isolated from fecal samples collect... Full description

Journal Title: FEMS microbiology letters 2006-04-01, Vol.257 (1), p.124-131
Main Author: Shima, Kensuke
Other Authors: Yoshii, Noriyo , Akiba, Masato , Nishimura, Kazuhiko , Nakazawa, Muneo , Yamasaki, Shinji
Format: Electronic Article Electronic Article
Language: English
Subjects:
Quelle: Alma/SFX Local Collection
Publisher: Oxford, UK: Blackwell Publishing Ltd
ID: ISSN: 0378-1097
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recordid: cdi_proquest_miscellaneous_67773066
title: Comparison of PCR-RFLP and PFGE for determining the clonality of enterohemorrhagic Escherichia coli strains
format: Article
creator:
  • Shima, Kensuke
  • Yoshii, Noriyo
  • Akiba, Masato
  • Nishimura, Kazuhiko
  • Nakazawa, Muneo
  • Yamasaki, Shinji
subjects:
  • Analysis
  • Animals
  • Assaying
  • Bacteria
  • Bacteriology
  • Biological and medical sciences
  • Cattle
  • Cattle Diseases - epidemiology
  • Cattle Diseases - microbiology
  • clonal turnover
  • Comparative analysis
  • E coli
  • Electrophoresis
  • Electrophoresis, Gel, Pulsed-Field - methods
  • Enterohemorrhagic E. Coli
  • Epidemiology
  • Escherichia coli
  • Escherichia coli Infections - epidemiology
  • Escherichia coli Infections - veterinary
  • Escherichia coli Infections - virology
  • Escherichia coli O157 - classification
  • Escherichia coli O157 - genetics
  • Escherichia coli O157 - isolation & purification
  • Feces - microbiology
  • Fundamental and applied biological sciences. Psychology
  • Gastrointestinal system
  • Gastrointestinal tract
  • Gel electrophoresis
  • Genetics
  • Microbiology
  • Molecular Epidemiology - methods
  • PCR‐restriction fragment length polymorphism
  • Phages
  • Polymerase chain reaction
  • Polymerase Chain Reaction - methods
  • Polymorphism
  • Polymorphism, Restriction Fragment Length
  • Pulsed-field gel electrophoresis
  • Restriction fragment length polymorphism
  • Shiga toxin
  • Stx‐phage
  • Systematics
ispartof: FEMS microbiology letters, 2006-04-01, Vol.257 (1), p.124-131
description: Abstract We report here on a comparative evaluation of PCR-restriction fragment length polymorphism (PCR-RFLP) and pulsed-field gel electrophoresis (PFGE) assays, and ascertain the clonal relationship between 13 enterohemorrhagic Escherichia coli O157 : H7 strains isolated from fecal samples collected from three cows over a period of 2 months. PCR-RFLP analysis was carried out with either BglI or EcoRV digested LA-PCR amplicons, generated by targeting region V of the Stx-phage. While PCR-RFLP analysis placed these 13 strains into a single clonal type, pulsotyping analysis, as reported earlier, grouped these strains into four different PFGE subtypes of which three were closely related, while the other appeared to be different. The comparative analysis was extended further using two clonally different wild-type (3-0 and Sakai 215) strains and 17 derivative strains which had passed through an animal's gastrointestinal tract. The PCR-RFLP assay, which was not only able to differentiate the wild-type strains, but also placed the passaged derivative strains into their respective parental group, although PFGE patterns of the same set of strains resulted from different PFGE subtypes. These data indicate that PCR-RFLP is the more reliable and useful assay for a molecular epidemiological survey of enterohemorrhagic E. coli strains.
language: eng
source: Alma/SFX Local Collection
identifier: ISSN: 0378-1097
fulltext: fulltext
issn:
  • 0378-1097
  • 1574-6968
url: Link


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titleComparison of PCR-RFLP and PFGE for determining the clonality of enterohemorrhagic Escherichia coli strains
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creatorShima, Kensuke ; Yoshii, Noriyo ; Akiba, Masato ; Nishimura, Kazuhiko ; Nakazawa, Muneo ; Yamasaki, Shinji
creatorcontribShima, Kensuke ; Yoshii, Noriyo ; Akiba, Masato ; Nishimura, Kazuhiko ; Nakazawa, Muneo ; Yamasaki, Shinji
descriptionAbstract We report here on a comparative evaluation of PCR-restriction fragment length polymorphism (PCR-RFLP) and pulsed-field gel electrophoresis (PFGE) assays, and ascertain the clonal relationship between 13 enterohemorrhagic Escherichia coli O157 : H7 strains isolated from fecal samples collected from three cows over a period of 2 months. PCR-RFLP analysis was carried out with either BglI or EcoRV digested LA-PCR amplicons, generated by targeting region V of the Stx-phage. While PCR-RFLP analysis placed these 13 strains into a single clonal type, pulsotyping analysis, as reported earlier, grouped these strains into four different PFGE subtypes of which three were closely related, while the other appeared to be different. The comparative analysis was extended further using two clonally different wild-type (3-0 and Sakai 215) strains and 17 derivative strains which had passed through an animal's gastrointestinal tract. The PCR-RFLP assay, which was not only able to differentiate the wild-type strains, but also placed the passaged derivative strains into their respective parental group, although PFGE patterns of the same set of strains resulted from different PFGE subtypes. These data indicate that PCR-RFLP is the more reliable and useful assay for a molecular epidemiological survey of enterohemorrhagic E. coli strains.
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subjectAnalysis ; Animals ; Assaying ; Bacteria ; Bacteriology ; Biological and medical sciences ; Cattle ; Cattle Diseases - epidemiology ; Cattle Diseases - microbiology ; clonal turnover ; Comparative analysis ; E coli ; Electrophoresis ; Electrophoresis, Gel, Pulsed-Field - methods ; Enterohemorrhagic E. Coli ; Epidemiology ; Escherichia coli ; Escherichia coli Infections - epidemiology ; Escherichia coli Infections - veterinary ; Escherichia coli Infections - virology ; Escherichia coli O157 - classification ; Escherichia coli O157 - genetics ; Escherichia coli O157 - isolation & purification ; Feces - microbiology ; Fundamental and applied biological sciences. Psychology ; Gastrointestinal system ; Gastrointestinal tract ; Gel electrophoresis ; Genetics ; Microbiology ; Molecular Epidemiology - methods ; PCR‐restriction fragment length polymorphism ; Phages ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Polymorphism ; Polymorphism, Restriction Fragment Length ; Pulsed-field gel electrophoresis ; Restriction fragment length polymorphism ; Shiga toxin ; Stx‐phage ; Systematics
ispartofFEMS microbiology letters, 2006-04-01, Vol.257 (1), p.124-131
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descriptionAbstract We report here on a comparative evaluation of PCR-restriction fragment length polymorphism (PCR-RFLP) and pulsed-field gel electrophoresis (PFGE) assays, and ascertain the clonal relationship between 13 enterohemorrhagic Escherichia coli O157 : H7 strains isolated from fecal samples collected from three cows over a period of 2 months. PCR-RFLP analysis was carried out with either BglI or EcoRV digested LA-PCR amplicons, generated by targeting region V of the Stx-phage. While PCR-RFLP analysis placed these 13 strains into a single clonal type, pulsotyping analysis, as reported earlier, grouped these strains into four different PFGE subtypes of which three were closely related, while the other appeared to be different. The comparative analysis was extended further using two clonally different wild-type (3-0 and Sakai 215) strains and 17 derivative strains which had passed through an animal's gastrointestinal tract. The PCR-RFLP assay, which was not only able to differentiate the wild-type strains, but also placed the passaged derivative strains into their respective parental group, although PFGE patterns of the same set of strains resulted from different PFGE subtypes. These data indicate that PCR-RFLP is the more reliable and useful assay for a molecular epidemiological survey of enterohemorrhagic E. coli strains.
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12Electrophoresis
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15Epidemiology
16Escherichia coli
17Escherichia coli Infections - epidemiology
18Escherichia coli Infections - veterinary
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20Escherichia coli O157 - classification
21Escherichia coli O157 - genetics
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23Feces - microbiology
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atitleComparison of PCR-RFLP and PFGE for determining the clonality of enterohemorrhagic Escherichia coli strains
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abstractAbstract We report here on a comparative evaluation of PCR-restriction fragment length polymorphism (PCR-RFLP) and pulsed-field gel electrophoresis (PFGE) assays, and ascertain the clonal relationship between 13 enterohemorrhagic Escherichia coli O157 : H7 strains isolated from fecal samples collected from three cows over a period of 2 months. PCR-RFLP analysis was carried out with either BglI or EcoRV digested LA-PCR amplicons, generated by targeting region V of the Stx-phage. While PCR-RFLP analysis placed these 13 strains into a single clonal type, pulsotyping analysis, as reported earlier, grouped these strains into four different PFGE subtypes of which three were closely related, while the other appeared to be different. The comparative analysis was extended further using two clonally different wild-type (3-0 and Sakai 215) strains and 17 derivative strains which had passed through an animal's gastrointestinal tract. The PCR-RFLP assay, which was not only able to differentiate the wild-type strains, but also placed the passaged derivative strains into their respective parental group, although PFGE patterns of the same set of strains resulted from different PFGE subtypes. These data indicate that PCR-RFLP is the more reliable and useful assay for a molecular epidemiological survey of enterohemorrhagic E. coli strains.
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