Development of a polymerase chain reaction assay for the rapid detection of the oral pathogenic bacterium, Selenomonas noxia.
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.
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.
Patricia
P
Cruz
MPH Program, Epidemiology & Biostatistics Concentration, Department of Environmental and Occupational Health, School of Community Health Sciences, University of Nevada, Las Vegas, NV, USA. mehretu@snhdmail.org.|Southern Nevada Health District, Las Vegas, NV, USA. mehretu@snhdmail.org.
Arthuro M
AM
Mehretu
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. mark.buttner@unlv.edu.
Mark P
MP
Buttner
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. theresa.trice@unlv.edu.
Theresa
T
Trice
Department of Biomedical Sciences, School of Dental Medicine, University of Nevada, Las Vegas, NV, USA. katherine.howard@unlv.edu.
Katherine M
KM
Howard
D001409:Bacillus cereus / Q000235:genetics; D002176:Candida albicans / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D006089:Gram-Negative Anaerobic Bacteria / Q000235:genetics; D006801:Humans; D007711:Klebsiella pneumoniae / Q000235:genetics; D007779:Lactobacillus acidophilus / Q000235:genetics; D009055:Mouth / Q000382:microbiology*; D009765:Obesity / Q000382:microbiology; D045852:Pectinatus / Q000235:genetics; D010510:Periodontal Diseases / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D015203:Reproducibility of Results; D020357:Selenomonas / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics; D013295:Streptococcus mutans / Q000235:genetics
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial