International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
권호 표지
DOI QR코드

DOI QR Code

Development of Species-specific PCR Primers for Detecting Peptoniphilus mikwangii

  • Park, Soon-Nang (Korean Collection for Oral Microbiology and Department of Oral Biochemistry, School of Dentistry, Chosun University) ;
  • Lee, Junhyeok (Farragut High School) ;
  • Kook, Joong-Ki (Korean Collection for Oral Microbiology and Department of Oral Biochemistry, School of Dentistry, Chosun University)
  • Received : 2017.08.10
  • Accepted : 2017.09.05
  • Published : 2017.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

In a previous study, Peptoniphilus mikwangii was isolated from the human oral cavity as a new species. The purpose of this study was to develop P. mikwangii-specific PCR primers. The PCR primers were designed, based on the nucleotide sequence of 16S ribosomal RNA (16S rDNA). The specificity of the primers was tested using genomic DNAs of 3 strains of P. mikwangii and 27 strains (27 species) of non-P. mikwangii bacteria. The sensitivity of primers sensitivity was determined using PCR, with serial dilutions of the purified genomic DNAs (4 ng to 4 fg) of P. mikwangii KCOM $1628^T$. The data showed that P. mikwangii-specific qPCR primers (B134-F11/B134-R1 & B134-F5/B134-R5) could detect only P. mikwangii strains, and 400 fg or 40 fg of P. mikwangii genome DNA. These results suggest that PCR primers are useful in detecting P. mikwangii from the oral cavity.

Keywords

References

  1. Cho E, Park SN, Shin Y, Lim YK, Paek J, Kim HK, Hwang CH, Jo E, Jin D, Chang YH, Kook JK. Peptoniphilus mikwangii sp. nov., isolated from a clinical specimen of human origin. Curr Microbiol. 2015;70:260-266. doi:10.1007/s00284-014-0712-7.
  2. Murdoch DC. Gram-positive anaerobic cocci. Clin Microbiol Rev. 1998;11:181-120.
  3. Ezaki T, Kawamura Y, Li N, Li ZY, Zhao L, Shu S. Proposal of the genera Anaerococcus gen. nov., Peptoniphilus gen. nov. and Gallicola gen. nov. for members of the genus Peptostreptococcus. Int J Syst Evol Microbiol. 2001;51:1521-1528. https://doi.org/10.1099/00207713-51-4-1521
  4. Murdoch DA, Shah HN. Reclassification of Peptostreptococcus magnus (Prevot 1933) Holdeman and Moore 1972 as Finegoldia magna comb. nov. and Peptostreptococcus micros (Prevot 1933) Smith 1957 as Micromonas micros comb. nov. Anaerobe 1999;5:555-559. https://doi.org/10.1006/anae.1999.0197
  5. Tindall BJ, Euzeby JP. Proposal of Parvimonas gen. nov. and Quatrionicoccus gen. nov. as replacements for the illegitimate, prokaryotic, generic names Micromonas Murdoch and Shah 2000 and Quadricoccus Maszenan et al. 2002, respectively. Int J Syst Evol Microbiol. 2006;56:2711-2713. https://doi.org/10.1099/ijs.0.64338-0
  6. Lee SM, Yoo SY, Kim HS, Kim KW, Yoon YJ, Lim SH, Shin HY, Kook JK. Prevalence of putative periodontopathogens in subgingival dental plaques from gingivitis lesions in Korean orthodontic patients. J Microbiol. 2005;43:260-265.
  7. Ashimoto A, Chen C, Bakker I, Slots J. Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions. Oral Microbiol Immunol. 1996;11:266-273. https://doi.org/10.1111/j.1399-302X.1996.tb00180.x
  8. Lee YJ, Kim MK, Hwang HK, Kook JK. Isolation and identification of bacteria from the root canal of the teeth diagnosed as the acute pulpitis and acute periapical abscess. J Korean Acad Conserv Dent. 2005;30:409-422. doi: https://doi.org/10.5395/JKACD.2005.30.5.409.
  9. Kim SG, Kim SH, Kim MK, Kim HS, Kook JK. Identification of Actinobacillus actinomycetemcomitans using speciesspecific 16S rDNA primers. J Microbiol. 2005;43:209-212.
  10. Han MJ, Chung CH, Kim HJ, Kook JK, Yoo SY. Detection of periodontal disease related bacteria from the implantabutment interface in oral cavity. J Korean Acad Prosthodont. 2008;46:116-124.
  11. Cho E, Park SN, Kim HK, Kim DS, Jung J, Baek JH, Lim YK, Jo E, Choi MH, Chang YH, Shin Y, Paek J, Shin JH, Kim J, Choi SH, Park HS, Kim H, Kook JK. Draft genome sequence of the novel Peptoniphilus sp. strain ChDC B134, isolated from a human periapical abscess lesion. Genome Announc. 2013;1:pii:e00822-13. doi: 10.1128/genomeA.00822-13.
  12. Shin HS, Kim MJ, Kim HS, Park SN, Kim do K, Baek DH, Kim C, Kook JK. Development of strain-specific PCR primers for the identification of Fusobacterium nucleatum subsp. fusiforme ATCC 51190T and subsp. vincentii ATCC 49256T. Anaerobe 2010;16:43-46. doi: 10.1016/j.anaerobe.2009.04.003.
  13. Krieg NR. Identification of procaryotes. In Boone DR, Castenholz RW, Garrity GM, editors. Bergeys manual of systematic bacteriology. 2nd ed. vol. 1. New York: Springer; 2001. p. 33-38.