Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Multiplex PCR Using Conserved and Species-Specific 16S rDNA Primers for Simultaneous Detection of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans

  • Kim, Mi-Kwang (Department of Oral Biochemistry, Chosun University, BK21, College of Dentistry, Chosun University) ;
  • Kim, Hwa-Sook (Department of Oral Biochemistry, Chosun University, BK21, College of Dentistry, Chosun University) ;
  • Kim, Byung-Ock (Oral Biology Research Institute, Chosun University) ;
  • Yoo, So-Young (Department of Oral Biochemistry, Chosun University, Oral Biology Research Institute, Chosun University) ;
  • Seong, Jin-Hyo (Oral Biology Research Institute, Chosun University) ;
  • Kim, Dong-Kie (Oral Biology Research Institute, Chosun University) ;
  • Lee, Shee-Eun (Department of Dental Pharmarcology, College of Dentistry, Chonnam National University) ;
  • Choe, Son-Jin (Department of Oral Microbiology and Immunology, Seoul National University) ;
  • Park, Joo-Cheol (Oral Biology Research Institute, Chosun University, BK21, College of Dentistry, Chosun University) ;
  • Min, Byung-Moo (Department of Oral Biochemistry, College of Dentistry, Seoul National University) ;
  • Jeong, Moon-Jin (Oral Biology Research Institute) ;
  • Kim, Do-Kyung (Oral Biology Research Institute) ;
  • Shin, Yong-Kook (Division of Genome Resources Bank and Reservation, National Genome Research Institute) ;
  • Kook, Joong-Ki (Department of Oral Biochemistry, Chosun University, Oral Biology Research Institute, Chosun University)
  • Published : 2004.02.01
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Abstract

This study was undertaken to develop PCR primers for the simultaneous detection of Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans, using two species-specific reverse primers in combination with a single conserved forward primer. These primers target the variable and conserved regions of the 16S rDNA. The primer specificity was tested against (i) four F. nucleatum and three A. actinomycetemcomitans strains and (ii) seven representatives of the different species of oral bacteria. The primer sensitivity was determined by testing serial dilutions of the purified genomic DNA of F. nucleatum and A. actinomycetemcomitans. The data indicate that species-specific amplicons could be obtained for all the F. nucleatum and A. actinomycetemcomitans strains tested, which were not found in the seven other species. The multiplex PCR could detect as little as 4 fg of chromosomal DNA of F. nucleatum and A. actinomycetemcomitans simultaneously. These findings suggest that these PCR primers are highly sensitive and are suitable for applications in epidemiological studies, diagnosis, and monitoring F. nucleatum and A. actinomycetemcomitans after the treatment of periodontitis.

Keywords

References

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