Identification of Campylobacter spp. and discrimination from Helicobacter and Arcobacter spp. by direct sequencing of PCR-amplified cpn60 sequences and comparison to cpnDB, a chaperonin reference sequence database.
A robust method for the identification of Campylobacter spp. based on direct sequencing of PCR-amplified partial cpn60 sequences and comparison of these to a reference database of cpn60 sequences is reported. A total of 53 Campylobacter isolates, representing 15 species, were identified and distinguished from phenotypically similar Helicobacter and Arcobacter strains. Pairwise cpn60 sequence identities between Campylobacter spp. ranged from 71 to 92 %, with most between 71 and 79 %, making discrimination of these species obvious. The method described overcomes limitations of existing PCR-based methods, which require time-consuming and complex post-amplification steps such as the cloning of amplification products. The results of this study demonstrate the potential for use of the reference chaperonin sequence database, cpnDB, as a tool for identification of bacterial isolates based on cpn60 sequences amplified with universal primers.
National Research Council Plant Biotechnology Institute, Saskatoon, Saskatchewan, Canada.
Janet E
JE
Hill
British Columbia Centre for Disease Control, 655W 12th Ave, Vancouver, BC, V5Z 4R4, Canada.
Ana
A
Paccagnella
British Columbia Centre for Disease Control, 655W 12th Ave, Vancouver, BC, V5Z 4R4, Canada.
Kee
K
Law
Bacteriology and Enteric Diseases Program, National Microbiology Laboratory, Winnipeg, Manitoba, Canada.
Pasquale L
PL
Melito
Bacteriology and Enteric Diseases Program, National Microbiology Laboratory, Winnipeg, Manitoba, Canada.
David L
DL
Woodward
Bacteriology and Enteric Diseases Program, National Microbiology Laboratory, Winnipeg, Manitoba, Canada.
Lawrence
L
Price
Amy H
AH
Leung
Bacteriology and Enteric Diseases Program, National Microbiology Laboratory, Winnipeg, Manitoba, Canada.
Lai-King
LK
Ng
National Research Council Plant Biotechnology Institute, Saskatoon, Saskatchewan, Canada.
Sean M
SM
Hemmingsen
British Columbia Centre for Disease Control, 655W 12th Ave, Vancouver, BC, V5Z 4R4, Canada.
Swee Han
SH
Goh
D020640:Arcobacter / Q000235:genetics* / Q000302:isolation & purification*; D002167:Campylobacter / Q000235:genetics* / Q000302:isolation & purification*; D018833:Chaperonins / Q000235:genetics*; D030541:Databases, Genetic; D016998:Helicobacter / Q000235:genetics* / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012015:Reference Standards; D013045:Species Specificity
D018833:Chaperonins