Species identification of Mycoplasma bovis and Mycoplasma agalactiae based on the uvrC genes by PCR.
The DNA repair genes uvrC from Mycoplasma bovis and Mycoplasma agalactiae type strains were cloned and their nucleotide sequences were established. These sequences were used to design polymerase chain reaction (PCR) primer pairs for M. bovis and M. agalactiae. Each primer pair amplified a 1-6 kb fragment of the uvrC gene in the respective species. The specificity of the primer pairs for the two species was demonstrated through the lack of cross-amplifications in heterologous PCR reactions and in reactions using DNA from other mycoplasma species. Subsequent restriction enzyme analysis of the amplified uvrC gene segments from type and field strains of M. bovis and M. agalactiae showed that the uvrC genes are well conserved in both species but differ significantly between the two species. The diagnostic PCR assay enabled unambiguous identification of M. bovis and M. agalactiae strains isolated from geographically diverse places, even in cases where 16S rRNA gene sequence analysis was unable to discriminate between the two species.
Institute for Veterinary Bacteriology, University of Berne, Switzerland.
S
S
Subramaniam
D
D
Bergonier
F
F
Poumarat
S
S
Capaul
Y
Y
Schlatter
J
J
Nicolet
J
J
Frey
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology; D003001:Cloning, Molecular; D017931:DNA Primers; D004706:Endodeoxyribonucleases; D029968:Escherichia coli Proteins; D015723:Gene Library; D005798:Genes, Bacterial; D015511:Goat Diseases / Q000175:diagnosis / Q000382:microbiology; D006041:Goats; D009174:Mycoplasma / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000382:microbiology; D013045:Species Specificity
D001426:Bacterial Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D004706:Endodeoxyribonucleases; C066567:UvrC protein, E coli