Development of a multiplex PCR assay for identification of Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp. hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis.
A multiplex PCR assay has been developed for the identification of the six common Campylobacter taxa associated with human gastroenteritis and/or septicaemia, namely Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp. hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis. The assay was developed using a combination of newly designed and published primers. It provided a specific PCR product for each of the five Campylobacter species and the one subspecies, and each of the PCR products was sufficiently distinguished by a difference in size by agarose gel electrophoresis. On evaluation of efficacy with 142 Campylobacter strains, the assay correctly identified all strains as 1 of the 6 Campylobacter taxa. This multiplex PCR assay is a rapid, simple and practical tool for identification of the six Campylobacter taxa commonly associated with gastroenteritis and/or septicaemia in humans, and offers an effective alternative to conventional biochemical-based assays.
Yamazaki-Matsune
Wataru
W
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Taguchi
Masumi
M
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Seto
Kazuko
K
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Kawahara
Ryuji
R
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Kawatsu
Kentaro
K
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Kumeda
Yuko
Y
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
Kitazato
Miyoshi
M
Yodogawa Christian Hospital, Osaka, Japan.
Nukina
Masafumi
M
Microbiological Department, Kobe Institute of Health, Hyogo, Japan.
Misawa
Naoaki
N
Department of Veterinary Public Health, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
Tsukamoto
Teizo
T
Division of Bacteriology, Osaka Prefectural Institute of Public Health, Osaka, Japan.
D002167:Campylobacter / Q000145:classification* / Q000235:genetics; D002169:Campylobacter Infections / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005759:Gastroenteritis / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D018805:Sepsis / Q000382:microbiology; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S