Detection and identification of Campylobacter spp. using the polymerase chain reaction.
Since Campylobacters have fastidious growth requirements and conventional detection and identification requires at least 4-6 days, the development of fast but reliable detection procedures is needed. Although methods based on DNA probe technology have been developed, these are not sensitive enough for the detection of Campylobacter spp. in food products. Therefore a PCR procedure based on the amplification of the 16S rRNA gene was developed that specifically detects the thermophilic Campylobacter species. This assay provides an excellent tool for the rapid and sensitive isolation and identification of Campylobacter spp. from chicken samples. In order to further identify the different Campylobacter spp., which are difficult to distinguish by conventional methods, PCR mediated approximately DNA typing was used to select species-specific DNA probes. This combination of PCR fingerprinting and probe hybridization results in a highly specific identification assay and provides an example of specific test development without the prior need for DNA sequence information. PCR mediated DNA typing was also used to study the epidemiology of diarrheal diseases caused by Campylobacter spp. Using primers complementary to dispersed repetitive DNA sequences and arbitrarily chosen DNA motifs PCR fingerprinting has proven to be a fast, highly discriminative and relatively simple method that can be applied in epidemiological investigations on Campylobacter infections. Besides this application of PCR fingerprinting for typing of Campylobacter spp. this method can also be used for the development of specific DNA probes.
Giesendorf
B A
BA
Academic Hospital Leiden, Department of Medical Microbiology, The Netherlands.
Quint
W G
WG
D000818:Animals; D015373:Bacterial Typing Techniques; D001431:Bacteriological Techniques; D001483:Base Sequence; D002167:Campylobacter / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D002169:Campylobacter Infections / Q000175:diagnosis / Q000382:microbiology; D016172:DNA Fingerprinting; D015342:DNA Probes; D005516:Food Microbiology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D015342:DNA Probes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S