Comparison of different techniques of quantitative PCR for determination of Streptococcus mutans counts in saliva samples.
Saliva samples from 16 children with current caries activity were investigated for Streptococcus mutans using three different PCR techniques, and the results were compared with those of selective cultivation on mitis salivarius agar with bacitracin (MSB) (I, II: LightCycler - competitive PCR end-point analysis; III: LightCycler - kinetic real-time analysis; IV, V: block cycler - competitive PCR end-point analysis; VI: cultivation on MSB agar). In groups I, III, IV and VI the saliva samples were analyzed directly. A DNA preparation before PCR with added competitors was carried out in groups II and V to exclude the influence of PCR inhibitors. The coefficients of correlation ranged from 0.97 to 0.98 among the competitive PCR methods, 0.8 to 0.85 for competitive vs. real-time PCR and 0.5 to 0.65 for PCR vs. cultivation methods. Competitive PCR on the real-time instrument was found to be more rapid than, comparably sensitive to, but less reproducible than competitive PCR on a block cycler.
Rupf
S
S
Department of Conservative Dentistry and Periodontology, University of Leipzig, Leipzig, Germany.
Merte
K
K
Kneist
S
S
Al-Robaiy
S
S
Eschrich
K
K
D001431:Bacteriological Techniques; D002648:Child; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis; D003731:Dental Caries / Q000382:microbiology*; D003732:Dental Caries Activity Tests; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012463:Saliva / Q000382:microbiology*; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric; D013295:Streptococcus mutans / Q000235:genetics / Q000302:isolation & purification*
D004269:DNA, Bacterial