International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
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Development of strain-specific polymerase chain reaction primers to detect Fusobacterium hwasookii strains

  • Lim, Yun Kyong (Korean Collection for Oral Microbiology and Department of Oral Biochemistry, School of Dentistry, Chosun University) ;
  • Kook, Joong-Ki (Korean Collection for Oral Microbiology and Department of Oral Biochemistry, School of Dentistry, Chosun University)
  • Received : 2021.10.19
  • Accepted : 2021.12.07
  • Published : 2021.12.29
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Abstract

This study aimed to develop strain-specific polymerase chain reaction (PCR) primers to detect Fusobacterium hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1256, F. hwasookii KCOM 1258, and F. hwasookii KCOM 1268 on the basis of nucleotide sequences of a gene specific to each strain. The unique genes for each F. hwasookii strain were determined on the basis of their genome sequences using Roary. The strain-specific PCR primers based on each strain-specific gene were designed using PrimerSelect. The specificity of each PCR primer was determined using the genomic DNA of the 5 F. hwasookii strains and 25 strains of oral bacterial species. The detection limit and sensitivity of each strain-specific PCR primer pair were determined using the genomic DNA of each target strain. The results showed that the strain-specific PCR primers correspond to F. hwasookii KCOM 1249T, F. hwasookii KCOM 1253, F. hwasookii KCOM 1258, F. hwasookii KCOM 1256/F. nucleatum subsp. polymorphum KCOM 1260, or F. hwasookii KCOM 1268/Fusobacterium sp. oral taxon 203 were developed. The detection limits of these strain-specific PCR primers ranged from 0.2 to 2 ng of genomic DNA for each target strain. The results suggest that these strain-specific PCR primers are valuable in quality control for detecting specific F. hwasookii strains.

Keywords

Acknowledgement

이 논문은 2020학년도 조선대학교 학술연구비(2020-737716-01)의 지원을 받아 연구되었음.

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