Polymerase chain reaction detection and speciation of Campylobacter upsaliensis and C. helveticus in human faeces and comparison with culture techniques.
A polymerase chain reaction (PCR) assay based on the 16S rRNA gene and an improved DNA extraction procedure were developed for the direct detection and differentiation of Campylobacter upsaliensis and C. helveticus in seeded human faeces. The PCR assay was compared with culture detection by a membrane filter (MF) technique and on selective agar (SA) containing 8 mg l-1 cefoperazone. Both MF culture and the PCR assay detected 10(5) colony-forming units (cfu) g-1 faeces. Selective agar culture of some strains could detect as few as 10(3) cfu g-1 faeces. However, some strains were susceptible to cefoperazone and either failed to grow or were detected only with reduced sensitivity in the presence of the antibiotic. Detection by MF and SA both required 48-96 h incubation in a microaerobic atmosphere and did not specifically identify the isolate. By contrast, the PCR assay could be completed within 8 h and accurately identified the two phenotypically similar species, C. upsaliensis and C. helveticus.
Lawson
A J
AJ
Laboratory of Enteric Pathogens, Central Public Health Laboratory, London, UK.
Linton
D
D
Stanley
J
J
Owen
R J
RJ
D000362:Agar; D002167:Campylobacter / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002482:Cellulose / Q000031:analogs & derivatives; D003470:Culture Media; D004269:DNA, Bacterial / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D006801:Humans; D008567:Membranes, Artificial; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012680:Sensitivity and Specificity
D003470:Culture Media; D004269:DNA, Bacterial; D008567:Membranes, Artificial; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; C005062:acetylcellulose; D000362:Agar; D002482:Cellulose