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Multiplex PCR using conserved and species-specific 16S rDNA primers for simultaneous detection of Fu

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Authors
Park, Joo Cheol; Kim, Mi-Kwang; Kim, Hwa Sook; Kim, Byung Ock; Yoo, So Young; Seong, Jin Hyo; Kim, Dong Kie; Lee, Shee Eun; Choe, Son Jin; Min, Byung Moo; Jeong, Moon Jin; Kim, Do Kyung; Shin, Yong Kook
Issue Date
2004
Publisher
한국미생물생명공학회
Citation
Journal of Microbiology and Biotechnology 14: 110-115
Keywords
actinobacillus actinomycetemcomitansfusobacterium nucleatummultiplex PCRrDNA
Abstract
this study was undertaken to develop PCR primers for the simultaneous detection of Fusobacterium nucleatum and Actinobacillus actinomyceltemcomitans, 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.
ISSN
1017-7825
Language
English
URI
https://hdl.handle.net/10371/66627
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College of Dentistry/School of Dentistry (치과대학/치의학대학원)Dept. of Dentistry (치의학과)Journal Papers (저널논문_치의학과)
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