Development of a combined filtration-enrichment culture followed by a one-step duplex PCR technique for the rapid detection of Campylobacter jejuni and C. coli in human faecal samples.
A new combined filtration-enrichment culture followed by a PCR technique for the rapid detection of Campylobacterjejuni and C. coli in human faeces has been developed. Only bacteria that passed through the membrane could multiply in the enrichment culture; target bacteria were detected by a one-step duplex PCR technique with combinations of primers that are specific for different Campylobacter spp., which should allow for the detection of a mixed infection in a single patient. A Falcon cell-culture insert and 24-well tissue-culture plates were used. After 2 days, both C. jejuni and C. coli were reliably detected in diluted faeces that were seeded with as few as 10 cells which corresponds to a concentration of 10(3) cfu/g. Even allowing for the dilution of faecal samples, this represents an increase in sensitivity of two-to-three orders of magnitude over the conventional filtration method.
Misawa
N
N
Department of Veterinary Public Health, Faculty of Agriculture, Miyazaki University, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
Kawashima
K
K
Department of Veterinary Public Health, Faculty of Agriculture, Miyazaki University, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
Kawamoto
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H
Department of Veterinary Public Health, Faculty of Agriculture, Miyazaki University, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
Kondo
F
F
Department of Veterinary Public Health, Faculty of Agriculture, Miyazaki University, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
D017000:Campylobacter coli / Q000235:genetics / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D003470:Culture Media; D017931:DNA Primers; D005243:Feces / Q000382:microbiology*; D005374:Filtration; D006801:Humans; D008851:Micropore Filters; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D003470:Culture Media; D017931:DNA Primers