PCR for detection and identification of Streptococcus sobrinus.
Oligonucleotide primers were designed based upon a comparison of the dextranase gene (dex) sequences from Streptococcus sobrinus and S. mutans. The primers amplified a 1610-bp long DNA fragment on the dex gene by a PCR. The pair of primers was specific to S. sobrinus as the other members of the mutans streptococci - S. mutans, S. downei, S. cricetus, S. rattus, S. macacae and S. ferus - gave no PCR products. Other gram-positive oral bacteria (15 strains of 10 species of cocci and 18 strains of 12 species of rods) and gram-negative oral bacteria (3 strains of 3 species of cocci and 31 strains of 22 species of rods) also gave negative results in the PCR. The PCR procedure was able to detect as little as 100 fg of purified chromosomal DNA or as few as 9 cfu of S. sobrinus NIDR6715. Seven clinical isolates of S. sobrinus were also positive in the dex PCR. This laboratory developed the S. mutans-specific PCR (dexA PCR) method with the primers specific for a portion of the dextranase gene of S. mutans Ingbritt. Primers for the dex and dexA PCR methods detected two species exclusively from the mutans streptococci. Furthermore, these two species were effectively differentiated by the species-specific amplicons with different lengths. The application of the PCR method to human dental plaque showed that the prevalence of S. sobrinus (83%) in oral cavities was higher than currently supposed (0-50%). These results suggest that the described PCR method is suitable for the specific detection and identification of human cariogenic bacteria, S. sobrinus and S. mutans.
Igarashi
T
T
Department of Oral Microbiology, Showa University School of Dentistry, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan.
Yamamoto
A
A
Department of Oral Microbiology, Showa University School of Dentistry, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan.
Goto
N
N
Department of Oral Microbiology, Showa University School of Dentistry, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142-8555, Japan.
D000818:Animals; D017931:DNA Primers / Q000235:genetics; D003773:Dental Plaque / Q000382:microbiology; D003910:Dextranase / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013290:Streptococcal Infections / Q000382:microbiology*; D013295:Streptococcus mutans / Q000145:classification / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification; D016986:Streptococcus sobrinus / Q000145:classification* / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D003910:Dextranase