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Development of a polymerase chain reaction assay for the rapid detection of the oral pathogenic bacterium, Selenomonas noxia.

Cruz P, Mehretu AM, Buttner MP, Trice T, Howard KM - BMC Oral Health (2015)

Bottom Line: The PCR assay was tested against fourteen non-target organisms, including closely related oral Selenomonads, one phylogenetically closely related bacterium, and two commonly isolated oral bacteria.One of the primer sets was more sensitive at detecting the target organism and was selected for optimization and validation experiments.The qPCR assay designed in this study can be used to specifically detect S. noxia in the clinical setting and in future research involving the enhanced detection of S. noxia.

View Article: PubMed Central - PubMed

Affiliation: Department of Environmental and Occupational Health, School of Community Health Sciences, University of Nevada Las Vegas, 4504 S. Maryland Parkway, Box 3064, Las Vegas, NV, 89154-3064, USA. patricia.cruz@unlv.edu.

ABSTRACT

Background: In recent studies, periodontal health has been linked to being overweight and/or obese. Among common oral bacteria, Selenomonas noxia has been implicated in converting periodontal health to disease, and Selenomonas species have also been found in gastric ulcers. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of S. noxia.

Methods: Two oligonucleotide primer pairs and one probe were designed and tested to determine optimal amplification signal with three strains of S. noxia. The PCR assay was tested against fourteen non-target organisms, including closely related oral Selenomonads, one phylogenetically closely related bacterium, and two commonly isolated oral bacteria.

Results: One of the primer sets was more sensitive at detecting the target organism and was selected for optimization and validation experiments. The designed primers and probe amplified the target organism with 100% specificity. PCR inhibition was observed with an internal positive control, and inhibition was resolved by diluting the DNA extract.

Conclusions: The qPCR assay designed in this study can be used to specifically detect S. noxia in the clinical setting and in future research involving the enhanced detection of S. noxia. The assay can also be used in epidemiological studies for understanding the role of S. noxia in disease processes including, but not limited to, oral health and obesity of infectious origin.

No MeSH data available.


Related in: MedlinePlus

Phylogenetic tree for the Selenomonads and related bacteria. The scale indicates a 5 % difference in nucleotide sequence, as determined by taking the sum of all branch lengths connecting two species [30]
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Fig1: Phylogenetic tree for the Selenomonads and related bacteria. The scale indicates a 5 % difference in nucleotide sequence, as determined by taking the sum of all branch lengths connecting two species [30]

Mentions: The phylogeny of species of Selenomonas and related bacteria has been determined by using 16S rRNA sequence analysis [24]. The Selenomonas group is phylogenetically coherent (Figure 1) with interspecies homology levels of 90 to 99 %. Selenomonas dianae, S. infelix, S. flueggei, S. noxia, S. artemidis, and Centipeda periodontii form a very tight cluster with a homology range of 96 to 99 %. Selenomonas sputigena and S. ruminantium have an average sequence homology of 94 % with members of this cluster [25].Figure 1


Development of a polymerase chain reaction assay for the rapid detection of the oral pathogenic bacterium, Selenomonas noxia.

Cruz P, Mehretu AM, Buttner MP, Trice T, Howard KM - BMC Oral Health (2015)

Phylogenetic tree for the Selenomonads and related bacteria. The scale indicates a 5 % difference in nucleotide sequence, as determined by taking the sum of all branch lengths connecting two species [30]
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4536781&req=5

Fig1: Phylogenetic tree for the Selenomonads and related bacteria. The scale indicates a 5 % difference in nucleotide sequence, as determined by taking the sum of all branch lengths connecting two species [30]
Mentions: The phylogeny of species of Selenomonas and related bacteria has been determined by using 16S rRNA sequence analysis [24]. The Selenomonas group is phylogenetically coherent (Figure 1) with interspecies homology levels of 90 to 99 %. Selenomonas dianae, S. infelix, S. flueggei, S. noxia, S. artemidis, and Centipeda periodontii form a very tight cluster with a homology range of 96 to 99 %. Selenomonas sputigena and S. ruminantium have an average sequence homology of 94 % with members of this cluster [25].Figure 1

Bottom Line: The PCR assay was tested against fourteen non-target organisms, including closely related oral Selenomonads, one phylogenetically closely related bacterium, and two commonly isolated oral bacteria.One of the primer sets was more sensitive at detecting the target organism and was selected for optimization and validation experiments.The qPCR assay designed in this study can be used to specifically detect S. noxia in the clinical setting and in future research involving the enhanced detection of S. noxia.

View Article: PubMed Central - PubMed

Affiliation: Department of Environmental and Occupational Health, School of Community Health Sciences, University of Nevada Las Vegas, 4504 S. Maryland Parkway, Box 3064, Las Vegas, NV, 89154-3064, USA. patricia.cruz@unlv.edu.

ABSTRACT

Background: In recent studies, periodontal health has been linked to being overweight and/or obese. Among common oral bacteria, Selenomonas noxia has been implicated in converting periodontal health to disease, and Selenomonas species have also been found in gastric ulcers. The objective of this study was to develop and validate a quantitative polymerase chain reaction (qPCR) assay for the specific and rapid detection of S. noxia.

Methods: Two oligonucleotide primer pairs and one probe were designed and tested to determine optimal amplification signal with three strains of S. noxia. The PCR assay was tested against fourteen non-target organisms, including closely related oral Selenomonads, one phylogenetically closely related bacterium, and two commonly isolated oral bacteria.

Results: One of the primer sets was more sensitive at detecting the target organism and was selected for optimization and validation experiments. The designed primers and probe amplified the target organism with 100% specificity. PCR inhibition was observed with an internal positive control, and inhibition was resolved by diluting the DNA extract.

Conclusions: The qPCR assay designed in this study can be used to specifically detect S. noxia in the clinical setting and in future research involving the enhanced detection of S. noxia. The assay can also be used in epidemiological studies for understanding the role of S. noxia in disease processes including, but not limited to, oral health and obesity of infectious origin.

No MeSH data available.


Related in: MedlinePlus