Typing of mutans streptococci by arbitrarily primed polymerase chain reaction.
The discriminative power of the arbitrarily primed polymerase chain reaction (AP-PCR) in differentiating between Streptococcus mutans and Strep. sobrinus species, serotypes and clones was investigated. Mutans streptococcal isolates (12(7)) obtained from 65 individuals (1-10 isolates per individual) were AP-PCR typed separately with two random primers, OPA-05 and OPA-13. Bacterial cell lysates were used as a template in PCR reactions, which made AP-PCR easy and fast to perform. Eighty-one isolates from 19 individuals were also ribotyped to compare the discriminative ability of ribotyping and AP-PCR techniques. AP-PCR performed with the two primers differentiated between Strep. mutans and Strep. sobrinus isolates, but neither primer detected serotype-specific amplification products. OPA-05 distinguished two main AP-PCR patterns among Strep. mutans isolates and one main pattern among Strep. sobrinus isolates, whereas OPA-13 found one main AP-PCR pattern among Strep. mutans isolates and two main patterns among Strep. sobrinus isolates. Ribotyping and AP-PCR revealed 40 and 33 different types among 81 selected isolates, respectively. Both techniques detected intra-individual heterogeneity in 16 out of 19 participants. The results indicate that AP-PCR has good discriminative ability in differentiating between mutans streptococcal clones and that the technique is suitable for epidemiological studies on mutans streptococci.
Saarela
M
M
Research Laboratory, University of Helsinki, Finland.
Hannula
J
J
Mättö
J
J
Asikainen
S
S
Alaluusua
S
S
D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D006801:Humans; D019105:Random Amplified Polymorphic DNA Technique; D015203:Reproducibility of Results; D012703:Serotyping; D013295:Streptococcus mutans / Q000145:classification* / Q000302:isolation & purification; D016986:Streptococcus sobrinus / Q000145:classification* / Q000302:isolation & purification
D004269:DNA, Bacterial; D004275:DNA, Ribosomal