Development of a 5′ Nuclease-Based Real-Time PCR Assay for Quantitative Detection of Cariogenic Dental Pathogens Streptococcus mutans and Streptococcus sobrinus

Abstract

A 5 nuclease TaqMan PCR assay was developed for the quantitative detection of the major cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus. The absolute and relative numbers of bacteria were measured by this method. This assay will be useful for quantifying these organisms in oral specimens and for analyzing biofilm formation.

Five-year longitudinal study of dental caries risk associated with <i>Streptococcus mutans</i> and <i>Streptococcus sobrinus</i> in individuals with intellectual disabilities

“…The primers and probes used in this study targeted the gtfB and gtfT genes of S. mutans and S. sobrinus, respectively, while another universal primer that targeted the eubacterial 16S rRNA gene was used to quantify the total bacterial load in the samples. The primers and probes (Table 1) were described previously (21,32).…”

“…In addition, various selective media have been used for isolation, quantification, and characterization of mutans streptococci (17). Previous studies indicate that polymerase chain reaction (PCR) methods are more sensitive for detection than conventional culture techniques (18)(19)(20), even though they are qualitative analytic techniques and therefore unsuitable for accurate evaluation of caries susceptibility or activity (21). Quantitative analysis is essential for monitoring cell numbers and the proportions of cariogenic bacteria in oral specimens such as dental plaque and saliva (21).…”

Regulatory change at Physalis Organ Size 1 correlates to natural variation in tomatillo reproductive organ size

“…Assays were performed with 1.0 ml cDNA, diluted two folds, using the SYBR Premix Ex Taq (TaKaRa). The quantification method (Delta-Delta CT) was used to evaluate variation between replicates 63 . The PFACTIN gene was used as an internal control to normalize the data.…”

Microbiological Diagnosis for Periodontal Diseases

“…Originally used for the measurement of DNA copy numbers, this technique has also been applied to the quantification of bacteria (Yoshida et al, 2003a;Yoshida et al, 2003b). One advantage of this technique is its wide dynamic range of bacterial detection, making it suitable for the determination of oral bacteria, which occur in various and variable amounts.…”

“…To date, many detection methods for bacteria in periodontal disease have been reported (Suzuki et al, 2004a;Suzuki et al, 2004b;Yoshida et al, 2003a;Yoshida et al, 2003b). Representative methods for the microbiological examination of periodontal disease are shown in Table 2.…”