Comparison of the value of pulsed-field gel electrophoresis, random amplified polymorphic DNA and amplified rDNA restriction analysis for subtyping Taylorella equigenitalis.
Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis.
Kagawa
S
S
Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, Japan.
Moore
J E
JE
Murayama
O
O
Matsuda
M
M
D000818:Animals; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods* / Q000662:veterinary; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary; D006734:Horse Diseases / Q000175:diagnosis / Q000382:microbiology; D006736:Horses; D016133:Polymerase Chain Reaction / Q000662:veterinary; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods* / Q000662:veterinary; D015183:Restriction Mapping / Q000662:veterinary; D020625:Taylorella equigenitalis / Q000737:chemistry / Q000145:classification* / Q000235:genetics
D004269:DNA, Bacterial; D004275:DNA, Ribosomal