Use of species-specific oligonucleotide probes to detect Mycoplasma gallisepticum, M. synoviae, and M. iowae PCR amplification products.
Three digoxigenin-labeled oligonucleotide probes, complementary to the variable region of the 16S ribosomal RNA (rRNA) gene of Mycoplasma gallisepticum, M. synoviae, and M. iowae were designed. The oligonucleotides were used in a dot blot hybridization assay. The target DNA is a 780-bp fragment of the 16S rRNA gene of avian mycoplasmas amplified by a single set of primers (multispecies polymerase chain reaction [PCR]). The oligonucleotide probes were specific for their corresponding PCR products at hybridization conditions of 56 C and 50% formamide. The detection limit of the dot blot hybridization assay was approximately 70, 50, and 30 colony-forming units for M. gallisepticum, M. synoviae, and M. iowae, respectively, per 4 microliters of PCR. In general, the oligonucleotide probe dot blotting assay was a more sensitive and effective method of detecting PCR products than detection by gel electrophoresis.
García
M
M
Department of Avian Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602-4875, USA.
Jackwood
M W
MW
Head
M
M
Levisohn
S
S
Kleven
S H
SH
D000818:Animals; D001483:Base Sequence; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000145:classification / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D014422:Turkeys
D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S