Polymerase chain reaction assay for detection of Campylobacter coli and Campylobacter jejuni in poultry meat.
Primers of 16S rRNA and flaA genes were used to optimise a PCR technique for detecting thermotolerant campylobacters in poultry meat. Different methods for crude DNA extraction were also evaluated. The use of flaA primers and extraction of nucleic acid by boiling and proteinase K gave good results in the detection of Campylobacter either in artificially or naturally contaminated foodstuffs. The lowest sensitivity limit for the PCR reaction was 10(1)-10(2) thermophilic Campylobacter cells either in pure cultures or in artificially and naturally contaminated poultry skins, corresponding to a concentration of 10(2)-10(3) Campylobacter/ml or g product. The PCR method we devised had a high sensitivity and specificity. It appears to give better results than conventional methods and is very easy and fast, requiring only eight hours to detect thermotolerant Campylobacter from poultry meat. In contrast, conventional methods require almost 4 days.
Manzano
M
M
Dipartimento di Scienze degli Alimenti, Università di Udine, Italy.
Pipan
C
C
Botta
G
G
Comi
G
G
D000818:Animals; D001483:Base Sequence; D017000:Campylobacter coli / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D016123:Campylobacter jejuni / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D005408:Flagellin / Q000235:genetics; D005798:Genes, Bacterial; D008460:Meat / Q000382:microbiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011200:Poultry; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D005408:Flagellin; C067138:flaA protein, bacteria