Specific PCR detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat.
An enrichment PCR assay using species-specific primers was developed for the detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. Primers for A. cryaerophilus, A. skirrowii, and A. cibarius were designed based on the gyrA gene to amplify nucleic acid fragments of 212, 257, and 145 bp, respectively. The A. butzleri-specific primers were designed flanking a 203-bp DNA fragment in the 16S rRNA gene. The specificity of the four primer pairs was assessed by PCR analysis of DNA from a panel of Arcobacter species, related Campylobacter, Helicobacter species, and other food bacteria. The applicability of the method was then validated by testing 42 fresh retail-purchased chicken samples in the PCR assay. An 18-h selective preenrichment step followed by PCR amplification with the four Arcobacter primer sets revealed the presence of Arcobacter spp. in 85.7% of the retail chicken samples analyzed. A. butzleri was the only species present in 50% of the samples, and 35.7% of the samples were positive for both A. butzleri and A. cryaerophilus. A. skirrowii and A. cibarius were not detected in any of the chicken samples analyzed. The enrichment PCR assay developed is a specific and rapid alternative for the survey of Arcobacter contamination in meat.
Pentimalli
Daniela
D
Dipartimento di Sanità Pubblica, Facoltà di Agraria, Università degli studi di Parma, Parma, Italy.
Pegels
Nicolette
N
García
Teresa
T
Martín
Rosario
R
González
Isabel
I
D000818:Animals; D020640:Arcobacter / Q000145:classification / Q000302:isolation & purification*; D001483:Base Sequence; D002645:Chickens; D015169:Colony Count, Microbial / Q000379:methods; D003470:Culture Media; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D005784:Gene Amplification; D006801:Humans; D008460:Meat / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000592:standards; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012680:Sensitivity and Specificity; D013045:Species Specificity
D003470:Culture Media; D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S