Immunocapture and real-time PCR to detect Campylobacter spp.
In this work, the feasibility of using a large-volume immunocapture system as a sample pretreatment before detection of Campylobacter was studied. Real-time PCR was used for detection of captured cells after immunocapture. This immunocapture system is able to process high-volume samples by recirculation, increasing the possibility of capturing cells in low numbers. After 30 min of recirculation, the sample is concentrated from 250 ml to 200 microl. In this study, different parameters were compared in order to improve cell capture. The analysis of inoculated chicken skin showed that detection of Campylobacter at levels of 10(3) CFU/25 g was possible after 8 h of enrichment. The low recovery of Campylobacter cells (< 1%) makes this separation method qualitative rather than quantitative. The detection limit of the entire protocol was increased due to the low cell recovery of the sample pretreatment. Therefore, this immunoseparation is able to recover cells present in high concentration after enrichment but not cells present in low concentration. Isolation of Campylobacter cells is achievable using this separation method rather than rapid detection.
Morales-Rayas
Rocío
R
Department of Food Science, University of Guelph, Guelph, Ontario, Canada N1G 2WI.
Wolffs
Petra F G
PF
Griffiths
Mansel W
MW
D000818:Animals; D002167:Campylobacter / Q000302:isolation & purification*; D002645:Chickens / Q000382:microbiology*; D015169:Colony Count, Microbial; D003257:Consumer Product Safety; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012867:Skin / Q000382:microbiology; D013997:Time Factors