Development and field validation of a Mycoplasma iowae real-time polymerase chain reaction assay.
A Mycoplasma iowae real-time polymerase chain reaction (PCR) assay using primers and probes targeting the 16S rRNA gene was developed and field-validated in this study. The assay specifically identified M. iowae with a detection limit of 80 colony-forming units (cfu) per turkey cloacal swab sample (3.2 cfu per PCR reaction). It was validated by testing 154 field turkey cloacal swab samples in parallel with culture isolation. The diagnostic sensitivity of the PCR was 97.6%, and the specificity was 95.5%. The real-time PCR developed in this study is a rapid, sensitive, and cost-effective alternative to culture isolation for detecting M. iowae from cloacal swab samples.
Cai
Hugh Y
HY
Animal Health Laboratory, Laboratory Services Division, University of Guelph, PO Box 3612, Guelph, Ontario, Canada N1H 6R8. hcai@lsd.uoguelph.ca
Bell-Rogers
Patricia
P
Parker
Lois
L
Ferencz
Arpad
A
Pozder
Peter
P
D000818:Animals; D001483:Base Sequence; D002988:Cloaca / Q000382:microbiology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D045793:Mycoplasma iowae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000175:diagnosis / Q000382:microbiology*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D012680:Sensitivity and Specificity; D014422:Turkeys
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S