DNA fingerprinting of isolates of Streptococcus mutans by pulsed-field gel electrophoresis.
Forty isolates and five standard laboratory strains, representing serotypes c, e and f of Streptococcus mutans were analyzed by pulsed-field gel electrophoresis (PFGE) after digestion of the genomic DNA with BssH II. The digestion patterns of standard laboratory strains were characteristic of serotypes c, e and f. Serotypes c and f generated diagnostic DNA fragments of approximately 145 kbp and of approximately 130-175 kbp in length, respectively. Serotype e generated a ladder of at least 14 fragments of 15-155 kbp in length. The digestion patterns of isolates were essentially similar to those of the standard laboratory strains. The patterns of almost all isolates obtained from a single individual were identical, but patterns of a few different types were also observed among isolates obtained from two individuals. Digestion with BssH II revealed differences among isolates obtained from different individuals. We used differences in banding patterns among isolates to construct a dendrogram. The dendrogram included two major clusters, one that consisted of isolates of serotypes c and f, and an other that consisted of isolates of serotype e. Our results indicate that BssH II is a useful enzyme for distinguishing among isolates of S. mutans and that digestion patterns obtained by PFGE can be used for chromosomal DNA fingerprinting.
Mineyama
R
R
Department of Oral Microbiology, School of Dentistry at Niigata, Nippon Dental University, 1-8 Hamauracho, 951-8580 Niigata, Japan. rmine@ecatv.home.ne.jp
Yoshino
S
S
Maeda
N
N
D015373:Bacterial Typing Techniques; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D006801:Humans; D015183:Restriction Mapping; D012463:Saliva / Q000382:microbiology; D013295:Streptococcus mutans / Q000145:classification* / Q000235:genetics
D004269:DNA, Bacterial; C044171:endodeoxyribonuclease BSSHII; D015252:Deoxyribonucleases, Type II Site-Specific