Genomics & Informatics

Korea Genome Organization (한국유전체학회)
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Effective microbial molecular diagnosis of periodontitis-related pathogen Porphyromonas gingivalis from salivary samples using rgpA gene

  • Jinuk Jeong (Department of Bioconvergence Engineering, Dankook University) ;
  • Yunseok Oh (Department of Bioconvergence Engineering, Dankook University) ;
  • Junhyeon Jeon (Department of Biological Sciences, Dankook University) ;
  • Dong-Heon Baek (Department of Oral Microbiology and Immunology, College of Dentistry, Dankook University) ;
  • Dong Hee Kim (Department of Anesthesiology and Pain Management, Dankook University Hospital) ;
  • Kornsorn Srikulnath (Animal Genomics and Bioresource Research Unit (AGB Research Unit), Faculty of Science, Kasetsart University) ;
  • Kyudong Han (Department of Bioconvergence Engineering, Dankook University)
  • Received : 2022.12.15
  • Accepted : 2023.01.06
  • Published : 2023.03.31
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Abstract

Importance of accurate molecular diagnosis and quantification of particular disease-related pathogenic microorganisms is highlighted as an introductory step to prevent and care for diseases. In this study, we designed a primer/probe set for quantitative real-time polymerase chain reaction (qRT-PCR) targeting rgpA gene, known as the specific virulence factor of periodontitis-related pathogenic bacteria 'Porphyromonas gingivalis', and evaluated its diagnostic efficiency by detecting and quantifying relative bacterial load of P. gingivalis within saliva samples collected from clinical subjects. As a result of qRT-PCR, we confirmed that relative bacterial load of P. gingivalis was detected and quantified within all samples of positive control and periodontitis groups. On the contrary, negative results were confirmed in both negative control and healthy groups. Additionally, as a result of comparison with next-generation sequencing (NGS)-based 16S metagenome profiling data, we confirmed relative bacterial load of P. gingivalis, which was not identified on bacterial classification table created through 16S microbiome analysis, in qRT-PCR results. It showed that an approach to quantifying specific microorganisms by applying qRT-PCR method could solve microbial misclassification issues at species level of an NGS-based 16S microbiome study. In this respect, we suggest that P. gingivalis-specific primer/probe set introduced in present study has efficient applicability in various oral healthcare industries, including periodontitis-related microbial molecular diagnosis field.

Keywords

Acknowledgement

The authors gratefully acknowledge Center for Bio-Medical Engineering Core Facility at Dankook University for providing critical reagents and equipment. The research institute has been supported by the VIP system as a part of the University Innovation Support Program 2022 of Dankook University. The research institute has been supported by the Department of Microbiology was supported through the Research-Focused Department Promotion & Interdisciplinary Convergence Research Project as a part of the Support Program for University Development for Dankook University in 2022.

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