Use of quantitative PCR to evaluate methods of bacteria sampling in periodontal patients

Masunaga, Hiroshi; Tsutae, Wataru; Oh, Hyun Ju; Shinozuka, Naoki; Kishimoto, Noriyoshi; Ogata, Yorimasa

doi:10.2334/josnusd.52.615

Cited by 21 publications

(12 citation statements)

References 21 publications

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“…Moreover, our results are in agreement with the findings of LauL et al 25 who found a clear increase in P. gingivalis and T. forsythensis was observed when comparing adult health to gingivitis and periodontitis patients. In addition, similar results were also reported by Kuboniwa et al 26 , Boutaga et al 27 , and Masunaga et al 28 . These results suggest that the number of major periodontopathogens increased with different periodontal disease status in adolescent.…”

Section: Discussionsupporting

confidence: 88%

Progression of periodontal inflammation in adolescents is associated with increased number of Porphyromonas gingivalis,Prevotella intermedia,Tannerella forsythensis, and Fusobacterium nucleatum

Yang

Zhang

et al. 2013

Int J Paed Dentistry

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Section: Discussionsupporting

confidence: 88%

Progression of periodontal inflammation in adolescents is associated with increased number of Porphyromonas gingivalis,Prevotella intermedia,Tannerella forsythensis, and Fusobacterium nucleatum

Yang

Zhang

et al. 2013

Int J Paed Dentistry

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“…In fact, all the strains which were not detected with the Invader assay had counts lower than 10 5 in real-time PCR. However, levels of periodontopathic bacterial species in subgingival plaque and saliva of patients with periodontitis ranged from 10 4 -10 7 copies (11,(21)(22)(23). These results suggest that the Invader assay is capable of detecting levels of periodontopathogens in patients with clinically problematic periodontal conditions.…”

Section: Discussionmentioning

confidence: 67%

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

et al. 2012

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SUMMARY: When quantifying periodontopathic bacteria, it is important to use a convenient method that does not produce false negative results. The Invader assay is a convenient method because it does not involve gene amplification. The purpose of this study was to evaluate the validity of the Invader assay to quantify periodontopathic bacteria. The Invader technology was applied in quantifying five periodontopathic bacteria (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, Prevotella intermedia, and Treponema denticola). The Invader assay produced a linear quantitative detection range over concentrations spanning seven exponential values, with a detection limit of 10 3.7 copies/tube and intra-day and inter-day variance of 0.1z to 4.7z and 0.1z to 3.4z, respectively, in quantifying five periodontopathic bacteria. We compared the results of the Invader assay with those of real-time polymerase chain reaction (PCR) performed for quantifying five periodontopathic bacteria in 22 patients with periodontitis. Among the Invader-detectable bacterial strains of each species, significant correlations were observed in the counts of concerned bacterial species between these two methods, with correlation coefficients ranging from 0.757 to 0.996. This study validated repeatability and reproducibility of the Invader assay in quantifying periodontopathic bacteria and demonstrated consistent agreement between the Invader assay and real-time PCR in quantifying periodontopathic bacteria.

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“…These assessments have been invaluable to dental basic science and translational research. Real‐time quantitative PCR (qPCR) has been used as an approach to rapidly quantify specific periodontal pathogens (Masunaga et al, 2010). In addition to real‐time qPCR, other DNA‐based techniques being used in assessing the oral bacteria include checkerboard hybridization (Dahlen et al, 2016; Goncalves et al, 2016; Vieira Colombo et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

Multiplex real‐time PCR detection and relative quantification of periodontal pathogens

Coffey

Choudhry

Shlossman

et al. 2016

Clinical & Exp Dental Res

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Periodontitis is a chronic inflammatory disease, which is strongly associated with certain pathogenic bacteria. The aim of this study was to develop a real‐time multiplex polymerase chain reaction (PCR) assay to detect and quantify bacterial species associated with periodontitis. We targeted detection and relative quantification of the following five bacterial species relevant to periodontal diseases: Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. The conserved regions of the genome of these species were targeted with oligos and TaqMan probes in real‐time PCR assays. The species‐specific TaqMan oligos and TaqMan probes showed no cross‐amplification, and there was no loss of amplification yield in multiplex real‐time PCR assays. All five bacterial targets were amplified analogous to the template concentrations used in these assays. This multiplex real‐time PCR strategy could potentially be used to detect the bacterial species in periodontal pockets of patients with periodontal diseases. This assay may also serve as a quick tool for profiling and quantifying bacteria relevant to periodontal diseases and likely be a valuable tool for clinical translational research.

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Use of quantitative PCR to evaluate methods of bacteria sampling in periodontal patients

Cited by 21 publications

References 21 publications

Progression of periodontal inflammation in adolescents is associated with increased number of Porphyromonas gingivalis,Prevotella intermedia,Tannerella forsythensis, and Fusobacterium nucleatum

Progression of periodontal inflammation in adolescents is associated with increased number of Porphyromonas gingivalis,Prevotella intermedia,Tannerella forsythensis, and Fusobacterium nucleatum

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

Multiplex real‐time PCR detection and relative quantification of periodontal pathogens

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