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.
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.
Wilson
Melissa K
MK
Department of Animal Sciences, University of Illinois, Urbana, IL, 61801, USA.
Lane
Alison B
AB
Law
Bibiana F
BF
Miller
William G
WG
Joens
Lynn A
LA
Konkel
Michael E
ME
White
Bryan A
BA
D000822:Animal Husbandry / Q000379:methods; D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods; D002169:Campylobacter Infections / Q000382:microbiology; D016123:Campylobacter jejuni / Q000145:classification / Q000235:genetics*; D002645:Chickens / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D014644:Genetic Variation; D016680:Genome, Bacterial; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial
Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):3140-5
9501229
J Clin Microbiol. 2005 May;43(5):2315-29
15872261
Nature. 2000 Feb 10;403(6770):665-8
10688204
Int J Food Microbiol. 2002 Apr 5;74(3):177-88
11981968
Nucleic Acids Res. 1991 Dec 25;19(24):6823-31
1762913
Emerg Infect Dis. 1999 Jan-Feb;5(1):28-35
10081669
Int J Syst Evol Microbiol. 2001 Jan;51(Pt 1):3-16
11211268
J Clin Microbiol. 2001 Jan;39(1):14-23
11136741
Appl Environ Microbiol. 2000 Jan;66(1):1-9
10618195
Appl Environ Microbiol. 2000 Sep;66(9):4029-36
10966425
Curr Opin Genet Dev. 2005 Dec;15(6):589-94
16185861
Nature. 2007 Oct 18;449(7164):804-10
17943116
Curr Microbiol. 2002 Aug;45(2):128-32
12070692
Emerg Infect Dis. 1999 Sep-Oct;5(5):607-25
10511517
Appl Environ Microbiol. 2003 Aug;69(8):4343-51
12902214
PLoS Biol. 2005 Jan;3(1):e15
15660156
Int J Food Microbiol. 2007 Jul 15;117(3):237-57
17368847
J Appl Microbiol. 1997 Dec;83(6):693-8
9449807