Development of strain-specific PCR primers for the identification of Fusobacterium nucleatum subsp. fusiforme ATCC 51190T and subsp. vincentii ATCC 49256T |
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| Authors: | Hwan Seon Shin Min-Jung Kim Hwa-Sook Kim Soon-Nang Park Do Kyung Kim Dong-Heon Baek Chan Kim Joong-Ki Kook |
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| Institution: | 1. Department of Oral Biochemistry, College of Dentistry, Chosun University, 375 Seo-Suk Dong, Dong-Ku, Gwangju 501-759, Republic of Korea;2. Department of Oral Physiology, College of Dentistry, Chosun University, 375 Seo-Suk Dong, Dong-Ku, Gwangju 501-759, Republic of Korea;3. The Second Stage of BK21 High-tech Dental Care and Human Resource Training Center, College of Dentistry, Chosun University, 375 Seo-Suk Dong, Dong-Ku, Gwangju 501-759, Republic of Korea;4. Department of Dental Hygiene, Chunnam Techno College, Gokseong County, Jeonnam 516-911, Republic of Korea;5. Department of Oral Microbiology and Immunology, College of Dentistry, Dankook University, Anseo-Dong, Cheonan, Chongnam 330-714, Republic of Korea;6. AMOMEDI Co. Ltd., 597-2 Wonsanri, Hasungmyun, Kimpo City, Kyungkido, Republic of Korea;7. AMOTECH Co. Ltd., New Materials Research Center, 597-2 Wonsanri, Hasungmyun, Kimpo City, Kyungkido 415-887, Republic of Korea;1. VCU-Shenandoah Family Practice Residency, Front Royal, VA, United States;2. Dickson Advanced Analytics, Carolinas HealthCare System, Charlotte, NC, United States;3. Department of Family Medicine, Carolinas Medical Center, Charlotte, NC, United States;1. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing High-tech Industrial Development Zone, Daqing 163319, PR China;2. Post-Doctoral Mobile Station of Veterinary Medicine, Northeast Agricultural University, No. 57 Mucai Street, Xiangfang District, Harbin 150030, PR China;1. The Lautenberg Center of General and Tumor Immunology, The Hebrew University Hadassah Medical School, IMRIC Jerusalem, 91120, Israel;2. The Rheumatology Research Center, Hadassah-Hebrew University, Jerusalem, 91120, Israel;3. The Institute of Dental Sciences, The Hebrew University-Hadassah School of Dental Medicine, Jerusalem, 91120, Israel;4. Department of General Surgery, Hadassah-Hebrew University Medical Center, Jerusalem, 91120, Israel;5. Department of Vascular Biology and Thrombosis Research Medical University of Vienna, 1090, Austria;6. Ella Institute of Melanoma, Sheba Medical Center, Ramat-Gan, 526260, Israel;7. Department of Histology and Embryology Center for Proteomics, Faculty of Medicine, University of Rijeka, 51000, Croatia;8. Harvard School of Public Health, Boston, MA, 02115, USA;9. Dana-Farber Cancer Institute, Boston, MA, 02115, USA;1. Department of Periodontology, Institute of Dentistry, University of Turku, Turku, Finland;2. Department of Biomedicine, Laboratory of Anatomy, University of Turku, Turku, Finland;1. Department of Surgery, University of Otago Christchurch, Christchurch, New Zealand;2. Microbiology Department, Canterbury Health Laboratories, Christchurch, New Zealand;3. Biostatistics and Computational Biology Unit, University of Otago Christchurch, Christchurch, New Zealand;1. Infectious Diseases Unit, Centre Hospitalier Marc Jacquet, 2, rue Fréteau de Pény, 77011 Melun, France;2. Intensive Care Unit, Centre Hospitalier Marc Jacquet, 2, rue Fréteau de Pény, 77011 Melun, France;3. Laboratoire de Bactériologie, Centre Hospitalier Marc Jacquet, 2, rue Fréteau de Pény, 77011 Melun, France |
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| Abstract: | The objective of this study was to develop the strain-specific PCR primers for Fusobacterium nucleatum subsp. fusiforme ATCC 51190T and F. nucleatum subsp. vincentii ATCC 49256T based on the nucleotide sequence of the Fs17 and Fv35 DNA probes, respectively. The strain specificity was tested against 10 type strains of Fusobacterium spp. or subsp., 21 clinical isolates of F. nucleatum from Koreans, and five type strains of distinct Fusobacterium species. Primer sensitivity was determined by testing serial dilutions (4 ng–4 fg) of the purified genomic DNA from each of the type strains. PCR showed that two pairs of PCR primers, Fs17-F14/Fs17-R14 and Fv35-F1/Fv35-R1 primers, could produce strain-specific amplicons from F. nucleatum subsp. fusiforme ATCC 51190T and F. nucleatum subsp. vincentii ATCC 49256T, respectively. The two PCR primer sets could detect as little as 0.4 pg or 4 pg of the genomic DNA of each target strain. These results suggest that the two sets of PCR primers could be used to identify F. nucleatum subsp. fusiforme ATCC 51190T and F. nucleatum subsp. vincentii ATCC 49256T, particularly for ascertaining the authenticity of the strain. |
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