International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
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Development of Streptococcus sanguinis-, Streptococcus parasanguinis-, and Streptococcus gordonii-PCR Primers Based on the Nucleotide Sequences of Species-specific DNA Probes Screened by Inverted Dot Blot Hybridization

  • Park, Soon-Nang (Korean Collection for Oral Microbiology, Department of Oral Biochemistry, and Oral Biology Research Institute, School of Dentistry, Chosun University) ;
  • Kook, Joong-Ki (Korean Collection for Oral Microbiology, Department of Oral Biochemistry, and Oral Biology Research Institute, School of Dentistry, Chosun University)
  • Received : 2013.02.20
  • Accepted : 2013.04.18
  • Published : 2013.06.30
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Abstract

The objective of this study was to develop PCR primers that are specific for Streptococcus sanguinis, Streptococcus parasanguinis, and Streptococcus gordonii. We designed the S. sanguinis-, S. parasanguinis-, and S. gordonii-specific primers, Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 respectively, based on the nucleotide sequences of the Ssa21, Spa17, and Sgo41 DNA probes that were screened using inverted dot blot hybridization (IDBH). The species-specificity of these primers was assessed against 43 strains of mitis group streptococci, including clinical strains of S. sanguinis, S. parasanguinis, and S. gordonii. The resulting PCR data revealed that species-specific amplicons had been obtained from all strains of the target species tested, and that none of these amplicons occurred in any other strains from other species. These results suggest that the Ssa21-F3/Ssa21-R2, Spa17-F/Spa17-R, and Sgo41-F1/Sgo41-R1 primers may be useful in detecting S. sanguinis, S. parasanguinis, and S. gordonii at the species level, respectively.

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References

  1. Kawamura Y, Hou XG, Sultana F, Miura H, Ezaki T. Determination of 16S rRNA sequences of Streptococcus mitis and Streptococcus gordonii and phylogenetic relationships among members of the genus Streptococcus. Int J Syst Bacteriol. 1995;45:406-408. https://doi.org/10.1099/00207713-45-2-406
  2. Gibbons RJ, Hay DI, Schlesinger DH. Delineation of a segment of adsorbed salivary acidic proline-rich proteins which promotes adhesion of Streptococcus gordonii to apatitic surfaces. Infect Immun. 1991;59:2948-2954.
  3. Scannapieco FA, Torres GI, Levine MJ. Salivary amylase promotes adhesion of oral streptococci to hydroxyapatite. J Dent Res. 1995;74:1360-1366. https://doi.org/10.1177/00220345950740070701
  4. Becker MR, Paster BJ, Leys EJ, Moeschberger ML, Kenyon SG, Galvin JL, Boches SK, Dewhirst FE, Griffen AL. Molecular analysis of bacterial species associated with childhood caries. J Clin Microbiol. 2002;40:1001-1009. https://doi.org/10.1128/JCM.40.3.1001-1009.2002
  5. Lamont RJ, Bevan CA, Gil S, Persson RE, Rosan B. Involvement of Porphyromonas gingivalis fimbriae in adherence to Streptococcus gordonii. Oral Microbiol Immunol. 1993; 8:272-276. https://doi.org/10.1111/j.1399-302X.1993.tb00573.x
  6. Caufield PW, Dasanayake AP, Li Y, Pan Y, Hsu J, Hardin JM. Natural history of Streptococcus sanguinis in the oral cavity of infants: evidence for a discrete window of infectivity. Infect Immun. 2000;68:4018-4023. https://doi.org/10.1128/IAI.68.7.4018-4023.2000
  7. Nyvad B, Kilian M. Comparison of the initial streptococcal microflora on dental enamel in caries-active and in cariesinactive individuals. Caries Res. 1990; 24:267-272. https://doi.org/10.1159/000261281
  8. Baddour LM. Virulence factors among gram-positive bacteria in experimental endocarditis. Infect. Immun. 1994; 62:2143-2148.
  9. Douglas CW, Heath J, Hampton KK, Preston FE. Identity of viridans streptococci isolated from cases of infective endocarditis. J Med Microbiol. 1993;39:179-182 https://doi.org/10.1099/00222615-39-3-179
  10. Herzberg MC, Gong K, MacFarlane GD, Erickson PR, Soberay AH, Krebsbach PH, Manjula G, Schilling K, Bowen WH. Phenotypic characterization of Streptococcus sanguis virulence factors associated with bacterial endocarditis. Infect Immun. 1990;58:515-522.
  11. 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;11:266-2673. https://doi.org/10.1111/j.1399-302X.1996.tb00180.x
  12. Conrads G, Flemmig TF, Seyfarth I, Lampert F, Lutticken R. Simultaneous detection of Bacteroides forsythus and Prevotella intermedia by 16S rRNA gene-directed multiplex PCR. J Clin Microbiol. 1999;37:1621-1624.
  13. Cho JS, Yoo SY, Kim HS, Hwang HK, Min JB, Kim BH, Baek DH, Shin HS, Kook JK. Identification and detection of Streptococcus anginosus using species-specific 16S rDNA primers. Int J Oral Biol. 2006;31:11-14.
  14. Kook JK, Kim MK, Seong JH, Kim DK, Kim BO, Park JC, Kim KK, Choe SJ, Min BM. A new method for rapid screening of bacterial species- or subspecies-specific DNA probes. FEMS Microbiol Lett. 2003;219:121-127. https://doi.org/10.1016/S0378-1097(03)00021-1
  15. Shin YK, Jeong SU, Yoo SY, Kim MK, Kim HS, Kim BO, Kim DK, Hwang HK, Kook JK. Pi30 DNA probe may be useful for the identification of Prevotella intermedia at the species or strain level. Microbiol Immunol. 2006;48: 931-936.
  16. Park CH, Kim PS, Kim HS, Min JB, Hwang HK, Jang HS, Cho KW, Baek DH, Kook JK. Detection of Prevotella intermedia and Prevotella nigrescens using Pn17 and Pn34 DNA Probes. Int J Oral Biol. 2010;35:13-19.
  17. Kim MJ, Hwang KH, Lee YS, Park JY, Kook JK. Development of Prevotella intermedia-specific PCR primers based on the nucleotide sequences of a DNA probe Pig27. J Microbiol Methods. 2011;84:394-397. https://doi.org/10.1016/j.mimet.2010.12.019
  18. Kim MJ, Lee YS, Park JY, Kook JK. Development of Prevotella nigrescens-specific PCR primers based on the nucleotide sequence of a Pn23 DNA probe. Anaerobe. 2011; 17:32-35. https://doi.org/10.1016/j.anaerobe.2010.12.005
  19. Kim HS, Song SK, Yoo SY, Jin DC, Shin HS, Lim CK, Kim MS, Kim JS, Choe SJ, Kook JK. Development of strain-specific PCR primers based on a DNA probe Fu12 for the identification of Fusobacterium nucleatum subsp. nucleatum ATCC $25586^T$. J Microbiol. 2005;43:331-336. Erratum in: J Microbiol. 2005;43:473.
  20. Shin HS, Kim MJ, Kim HS, Park SN, Kim DK, 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 $49256^T$. Anaerobe. 2010;16:43-46. https://doi.org/10.1016/j.anaerobe.2009.04.003
  21. Kim MJ, Min JB, Lim SA, Kook JK. Strain-specific PCR Primers for the Detection of Prevotella intermedia ATCC 49046. Int J Oral Biol. 2011;36:79-82.
  22. Yoo SY, Kim KJ, Lim SH, Kim KW, Hwang HK, Min BM, Choe SJ, Kook JK. First isolation of Streptococcus downei from human dental plaques. FEMS Microbiol Lett. 2005; 249:323-326. https://doi.org/10.1016/j.femsle.2005.06.020
  23. Rychlik W. Selection of primers for polymerase chain rea-- ction. 1st ed. In B.A. White (ed.), PCR protocols, Humana Press, Totowa, New Jersey, USA. 1993;31-40.
  24. Song SK, Yoo SY, Kim MK, Kim HS, Lim SA, Kim DK, Park JY, Kook JK. Development of Prevotella nigrescens ATCC $33563^T$-specific PCR primers. Kor J Microbiol. 2008;44: 221-227.