Rapid and simple detection of eight major periodontal pathogens by the loop-mediated isothermal amplification method

Junko Miyagawa, Hiroshi Maeda, Toshimitsu Murauchi, Susumu Kokeguchi, Kokoro Yamabe, Ichiro Tanimoto, Fusanori Nishimura, Kazuhiro Fukui, Shogo Takashiba

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Loop-mediated isothermal amplification (LAMP) was applied to develop a rapid and simple detection system for eight periodontal pathogens: Aggregatibacter (Actinobacillus) actinomycetemcomitans, Campylobacter rectus, Eikenella corrodens, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Tannerella forsythia. Primers were designed from the 16S ribosomal RNA gene for each pathogen, and the LAMP amplified the targets specifically and efficiently under isothermal condition at 64°C. To simplify the manipulation of LAMP examination, boiled cells and intact cells suspended in phosphate-buffered saline (PBS) were tested as templates besides extracted DNA template. The detection limits were 1-10 cells per tube using extracted DNA template. However, LAMP methods using boiled cells and intact cells required 10-100 and 100-1000 cells per tube, respectively. LAMPs for A. actinomycetemcomitans, P. gingivalis and P. intermedia were then applied to clinical plaque samples, and the method demonstrated equal or higher sensitivity compared with the conventional real-time PCR method. These findings suggest the usefulness of the LAMP method for the rapid and simple microbiological diagnosis of periodontitis, and the possibility of LAMP examination without the DNA extraction step.

Original languageEnglish
Pages (from-to)314-321
Number of pages8
JournalFEMS Immunology and Medical Microbiology
Volume53
Issue number3
DOIs
Publication statusPublished - Aug 1 2008

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

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