2 research outputs found
A polymerase chain reaction method for the detection of Selenomonas noxia
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. Selenomonas spp. have also been found in gastric ulcers, and were misdiagnosed as Helicobacter -like organisms, but no further studies were conducted. The increasing clinical and epidemiological importance of S. noxia necessitates the development of a rapid detection method. In this study, a TaqMan 16S rRNA based real-time Polymerase Chain Reaction (PCR) method was developed, optimized and evaluated for the rapid and specific detection of S. noxia . The 16S PCR assay using primers and a fluorescent probe was tested against S. noxia , six organisms closely related to S. noxia , and two commonly isolated oral bacteria. The designed primers and probe were optimized, and amplified the target organism with 100% specificity. In conclusion, the 16S real-time PCR designed in this study can be used to specifically amplify S. noxia . The assay can be used for epidemiological studies in understanding the role of S. noxia in disease processes including, but not limited to, oral health and infectobesity
Development of a New 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. noxiain 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