Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulsed-Field Gel Electrophoresis, Multilocus Sequence Typing (MLST), and Repetitive Sequence Polymerase Chain Reaction (rep-PCR) Reveals Different Discriminatory Capabilities

Melissa K. Wilson, Alison B. Lane, Bibiana Law, William G. Miller, Lynn A. Joens, Michael E. Konkel, Bryan A. White

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Campylobacter jejuni is one of the leading bacterial causes of food-borne illness in the USA. Molecular typing methods are often used in food safety for identifying sources of infection and pathways of transmission. Moreover, the identification of genetically related isolates (i.e., clades) may facilitate the development of intervention strategies for control and prevention of food-borne diseases. We analyzed the pan genome (i.e., core and variable genes) of 63 C. jejuni isolates recovered from chickens raised in conventional, organic, and free-range poultry flocks to gain insight into the genetic diversity of C. jejuni isolates recovered from different environments. We assessed the discriminatory power of three genotyping methods [i.e., pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and repetitive extragenic palindromic polymerase chain reaction (rep-PCR)]. The rep-PCR fingerprint was generated by determining the presence of repetitive sequences that are interspersed throughout the genome via repetitive extragenic palindromic PCR, enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR), and BOX element PCR (BOX-PCR) and combining the data to form a composite fingerprint. The genetic fingerprints were subjected to computer-assisted pattern analysis. Comparison of the three genotypic methods revealed that repREB-PCR showed greater discriminatory power than PFGE and MLST. ERIC-PCR and BOX-PCR yielded the highest number of PCR products and greatest reproducibility. Regardless of the genotyping method, C. jejuni isolates recovered from chickens reared in conventional, organic, and free-range environments all exhibit a high level of genotypic diversity.

Original languageEnglish (US)
Pages (from-to)843-855
Number of pages13
JournalMicrobial Ecology
Volume58
Issue number4
DOIs
StatePublished - Nov 2009

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repetitive sequences
Campylobacter jejuni
pulsed-field gel electrophoresis
poultry
polymerase chain reaction
electrokinesis
gel
genome
consensus sequence
foodborne illness
genotyping
free range husbandry
chickens
genetic variation
food
food safety
methodology
reproducibility
flocks
multilocus sequence typing

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Ecology
  • Soil Science

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Analysis of the Pan Genome of Campylobacter jejuni Isolates Recovered from Poultry by Pulsed-Field Gel Electrophoresis, Multilocus Sequence Typing (MLST), and Repetitive Sequence Polymerase Chain Reaction (rep-PCR) Reveals Different Discriminatory Capabilities. / Wilson, Melissa K.; Lane, Alison B.; Law, Bibiana; Miller, William G.; Joens, Lynn A.; Konkel, Michael E.; White, Bryan A.

In: Microbial Ecology, Vol. 58, No. 4, 11.2009, p. 843-855.

Research output: Contribution to journalArticle

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