Rapid PCR using nested primers of the 16S rRNA and the hippuricase (hip O) genes to detect Campylobacter jejuni and Campylobacter coli in environmental samples.
Identification of sources Campylobacter infection in the poultry houses is in general problematic due to the lack of reliable methods to detect campylobacteria in environmental samples. Detection of campylobacteria in environmental samples by conventional culture methods is difficult and of limited sensitivity due to the use of selective media, the low number of bacteria in the samples and possibly also due to the presence of non-culturable or sub-lethally injured stages of the bacteria. The present paper describes a rapid PCR assay using nested primers of the 16S rRNA or the hippuricase (hip O) genes to detect Campylobacter jejuni and Campylobacter coli in environmental samples. The sensitivity of the nested PCR was determined to be 0.01 pg/PCR, corresponding to 2-3 colony forming units (cfu) per ml. The nested PCR assays were applied to detect C. jejuni and C. coli in 269 environmental samples collected from ten broiler farms. The sensitivity, specificity and the usefulness of the PCR assay for detection of C. jejuni and C. coli in environmental samples are presented and discussed.
Bang
D D
DD
Department of Poultry, Fish, and Fur Animals, Danish Veterinary Institute, Hangøvej 2, DK-8200 Aarhus N, Denmark. ddb@vetinst.dk
Wedderkopp
A
A
Pedersen
K
K
Madsen
M
M
D000581:Amidohydrolases / Q000235:genetics*; D000818:Animals; D017000:Campylobacter coli / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000302:isolation & purification*; D002645:Chickens; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004777:Environment; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011201:Poultry Diseases / Q000382:microbiology; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity; D013997:Time Factors
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D000581:Amidohydrolases; C019672:aminoacylase I