Genetic characterization of the oral Actinomyces.
Actinomyces are difficult to identify using serological and biochemical methods but genotyping is an efficient and reliable means of bacterial characterization and can be used to determine clonal identity. The purpose here was to genotype 13 American type culture collection (ATCC) reference strains representing six different oral Actinomyces spp. by using chromosomal DNA fingerprinting (CDF), arbitrarily primed-polymerase chain reaction (AP-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In CDF analysis, BamHI, BstEII and SmaI yielded digestion patterns revealing characteristic differences among the known Actinomyces spp., with SmaI demonstrating optimal resolution. Amplicons generated by AP-PCR with primer OPB-07 displayed banding patterns that permitted discrimination of all Actinomyces strains tested. PCR-RFLP with MnlI digests generated fragment patterns that also characterized the reference strains. Collectively, genotypic profiles generated by CDF, AP-PCR and PCR-RFLP permitted differentiation of all 13 ATCC Actinomyces strains. SmaI CDF analysis of 18 clinical isolates of catalase-positive A. naeslundii genospecies 2 revealed extensive genetic diversity among these strains. These molecular approaches may be useful in determining genetic diversity within oral Actinomyces populations and fidelity of Actinomyces transmission between mother and child.
Ruby
J D
JD
Department of Oral Biology, School of Dentistry, The University of Alabama at Birmingham, SOD Box 13, 1919 Seventh Avenue South, Birmingham, AL 35294-0007, USA. john_ruby@csl.dental.uab.edu
Li
Y
Y
Luo
Y
Y
Caufield
P W
PW
D000190:Actinomyces / Q000145:classification / Q000235:genetics*; D002374:Catalase / Q000235:genetics; D002675:Child, Preschool; D002876:Chromosomes, Bacterial / Q000235:genetics; D016172:DNA Fingerprinting; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000235:genetics; D015245:Deoxyribonuclease BamHI; D015252:Deoxyribonucleases, Type II Site-Specific; D005260:Female; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D009055:Mouth / Q000382:microbiology*; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012463:Saliva / Q000382:microbiology
D004269:DNA, Bacterial; D002374:Catalase; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific