- Development of Strain-specific PCR Primers Based on a DNA Probe Fu12 for the Identification of Fusobacterium nucleatum subsp. nucleatum ATCC 25586^T
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Hwa-Sook Kim , Soo Keun Song , So Young Yoo , Dong Chun Jin , Hwan Seon Shin , Chae Kwang Lim , Myung-Soo Kim , Jin-Soo Kim , Son-Jin Choe , Joong-Ki Kook
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J. Microbiol. 2005;43(4):331-336.
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DOI: https://doi.org/2257 [pii]
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 Abstract
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The objective of this study was to assess the strain-specificity of a DNA probe, Fu12, for Fusobacterium nucleatum subsp. nucleatum ATCC 25586^T (F. nucleatum ATCC 25586^T), and to develop sets of strain-specific polymerase chain reaction (PCR) primers. Strain-specificity was tested against 16 strains of F. nucleatum and 3 strains of distinct Fusobacterium species. Southern blot hybridization revealed that the Fu12 reacted exclusively with the HindIII-digested genomic DNA of F. nucleatum ATCC 25586^T. The results of PCR revealed that three pairs of PCR primers, based on the nucleotide sequence of Fu12, generated the strain-specific amplicons from F. nucleatum ATCC 25586^T. These results suggest that the DNA probe Fu12 and the three pairs of PCR primers could be useful in the identification of F. nucleatum ATCC 25586^T, especially with regard to the determination of the authenticity of the strain.
- Prevalence of Putative Periodontopathogens in Subgingival Dental Plaques from Gingivitis Lesions in Korean Orthodontic Patients
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Seung Mi Lee , So Young Yoo , Hwa-Sook Kim , Kwang-Won Kim , Young-Joo Yoon , Sung-Hoon Lim , Hee-Young Shin , Joong-Ki Kook
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J. Microbiol. 2005;43(3):260-265.
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DOI: https://doi.org/2215 [pii]
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Abstract
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The objective of this study was to detect and compare the presence of periodontopathogens in the subgingival plaques of gingivitis lesions in adults who wore fixed orthodontic appliances, as opposed to adults who did not wear any orthodontic appliances. Thirty-six individuals participated in this study. Ninteen of these subjects did not wear any orthodontic appliances, and these subjects comprised the control group. The other 17 individuals had been wearing fixed orthodontic appliances for at least 3 months each. After a periodontal examination, we collected subgingival plaque samples from the gingivitis lesions of each patient. Using PCR based on 16S rDNA, we detected the presence of 6 putative periodontopathogenic species, Treponema denticola, Porphyromonas gingivalis, Tannerella forsythia (formerly Bacteroides forsythus), Prevotella nigrescens, Prevotella intermedia, and Actinobacillus actinomycetemcomitans. With regard to the presence of individual periodontopathogens, we found that T. forsythia, T. denticola, and P. nigrescens were significantly more common in the samples obtained from the orthodontic patients than in the samples obtained from the non-orthodontic patient controls. Our results indicate that the local changes associated with the wearing of fixed orthodontic appliances may affect the prevalence of periodontopathogens in subgingival dental plaques.
- Identification of Actinobacillus actinomycetemcomitans Using Species-Specific 16S rDNA Primers
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Su-Gwan Kim , Soo-Heung Kim , Mi-Kwang Kim , Hwa-Sook Kim , Joong-Ki Kook
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J. Microbiol. 2005;43(2):209-212.
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DOI: https://doi.org/2159 [pii]
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Abstract
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The purpose of this study was to develop species-specific PCR primers for use in the identification and detection of Actinobacillus actinomycetemcomitans. These primers target variable regions of the 16S ribosomal RNA coding gene (rDNA). We assessed the specificity of the primers against 9 A. actinomycetemcomitans strains and 11 strains (3 species) of the Haemophilus genus. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC 33384^T. Our obtained data revealed that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 4 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these PCR primers are incredibly sensitive, and should prove suitable for application in epidemiological studies, as well as the diagnosis and monitoring of periodontal pathogens after treatment for periodontitis.
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