Specific detection of Mycoplasma iowae using polymerase chain reaction.
A polymerase chain reaction assay was developed for the specific detection of Mycoplasma iowae, a poultry pathogen. Two primers were chosen in the DNA region located immediately 5' of the ribosomal genes operon. Under our PCR conditions, specific amplification of all M. iowae strains tested was achieved, and none of 17 other Mycoplasma species isolated from birds gave an amplification product. We obtained amplification from culture medium samples of M. iowae by using Tth DNA polymerase instead of Taq DNA polymerase, and we were able to detect as few as 10 organisms. Finally, we described a dot blot hybridization test, using cold labelling and chemiluminescence, which is very convenient for routine detection of M. iowae.
Laigret
F
F
Laboratoire de Biologie Cellulaire et Moléculaire, INRA et Université de Bordeaux II, Villenave d'Ornon, France.
Deaville
J
J
Bové
J M
JM
Bradbury
J M
JM
D000818:Animals; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D008163:Luminescent Measurements; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis; D012680:Sensitivity and Specificity; D013045:Species Specificity; D008973:Tenericutes / Q000235:genetics; D014422:Turkeys
D004269:DNA, Bacterial; D004275:DNA, Ribosomal