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Rapid Detection of Serratia fonticola by TaqMan Quantitative Real-Time PCR Using Primers Targeting the gyrB Gene

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Abstract

A gyrB gene is present in the majority of bacterial species, and encodes the ATPase domain of DNA gyraseB-subunit protein, which is essential for transcription and replication of bacteria. The gyrB gene exhibits higher nucleotide sequence variability than the 16S rDNA gene and thus could be more reliable in differentiating Serratia fonticola. A species-specific primer pair and probe were designed for quantitative real-time PCR detection of S. fonticola using gyrB as the target gene. Nine members of the Serratia family (representing nine Serratia species) were chosen to verify the specificity of the primers. Additionally, two species each of Salmonella and Klebsiella, and five other species belonging to five other genera of Enterobacteriaceae, were tested for primer cross-reaction. All the tested strains gave negative results. The limit of detection for S. fonticola using the gyrB gene was 100 copies per PCR reaction. This TaqMan PCR assay provided a specific, rapid, and sensitive method to detect S. fonticola based on its gyrB gene.

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References

  1. Aiyar A (2000) The use of CLUSTAL W and CLUSTAL X for multiple sequence alignment. Methods Mol Biol 132(132):221–241

    CAS  PubMed  Google Scholar 

  2. Aljorayid A, Viau R, Castellino L, Jump RLP (2016) Serratia fonticola, pathogen or bystander? A case series and review of the literature. IDCases 5:6–8

    Article  PubMed  PubMed Central  Google Scholar 

  3. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25(17):3389–3402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Bollet C, Gainnier M, Sainty JM, Orhesser P, Micco PD (1991) Serratia fonticola isolated from a leg abscess. J Clin Microbiol 29(4):834–835

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Farmer JJI, Davis BR (1985) Biochemical identification of new species and biogroups of Enterobacteriaceae isolated from clinical specimens. J Clin Microbiol 21(1):46

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Garcia ME, Lanzarot P, Costas E, Rodas VL, Marín M, Blanco JL (2008) Isolation of Serratia fonticola from skin lesions in a Nile Crocodile (Crocodylus niloticus) with an associated septicaemia. Vet J 176(2):254–256

    Article  CAS  PubMed  Google Scholar 

  7. Gavini F, Ferragut C, Izard D, Trinel PA, Leclerc H, Lefebvre B, Mossel DAA (1979) Serratia fonticola, a new species from water. Int J Syst Bacteriol 29(2):92–101

    Article  Google Scholar 

  8. Gellert M, Mizuuchi K, O’Dea MH, Nash HA (1976) DNA gyrase: an enzyme that introduces superhelical turns into DNA. Proc Natl Acad Sci USA 73(11):3872–3876

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Gorret J, Chevalier J, Gaschet A, Fraisse B, Violas P, Chapuis M, Jolivet-Gougeon A (2009) Childhood delayed septic arthritis of the knee caused by Serratia fonticola. Knee 16(6):512–514

    Article  PubMed  Google Scholar 

  10. Iwaya A, Nakagawa S, Iwakura N, Taneike I, Kurihara M, Kuwano T, Gondaira F, Endo M, Hatakeyama K, Yamamoto T (2005) Rapid and quantitative detection of blood Serratia marcescens by a real-time PCR assay: its clinical application and evaluation in a mouse infection model. FEMS Microbiol Lett 248(2):163–170

    Article  CAS  PubMed  Google Scholar 

  11. Jongstra J, Jongstra-Bilen J, Tidmarsh GF, Davis MM (2014) Use of quantitative real-time PCR for direct detection of Serratia marcescens in marine and other aquatic environments. Appl Environ Microbiol 80(5):1679–1683

    Article  Google Scholar 

  12. Kämpfer P, Glaeser SP (2015) Serratia glossinae Geiger et al. 2010 is a later synonym of Serratia fonticola Gavini et al. 1979. Int J Syst Evol Microbiol 65(Pt 5):1406–1408

    Article  PubMed  Google Scholar 

  13. Kasai H, Watanabe K, Gasteiger E, Bairoch A, Isono K, Yamamoto S, Harayama S (1998) Construction of the gyrB database for the identification and classification of bacteria. Genome Inf 9(4):13–21

    CAS  Google Scholar 

  14. Kunimoto D, Rennie R, Citron DM, Goldstein EJ (2004) Bacteriology of a bear bite wound to a human: case report. J Clin Microbiol 42(7):3374

    Article  PubMed  PubMed Central  Google Scholar 

  15. Mahlen SD (2011) Serratia infections: from military experiments to current practice. Clin Microbiol Rev 24(4):755–791

    Article  PubMed  PubMed Central  Google Scholar 

  16. Merino LA, Hreñuk GE, Alonso JM, Ronconi MC (2000) Usefulness of antibiotype and bacteriocinotype for differentiating Shigella strains isolated in Argentina. Bull Soc Pathol Exot 93(5):307

    CAS  PubMed  Google Scholar 

  17. Müller HE, Steigerwalt AG, Brenner DJ (1986) Isolation of Serratia fonticola from birds. Zbl Bakt Hyg A 261(2):212–218

    Google Scholar 

  18. Ochieng JB, Boisen N, Lindsay B, Santiago A, Ouma C, Ombok M, Fields B, Stine OC, Nataro JP (2014) Serratia marcescens is injurious to intestinal epithelial cells. Gut Microbes 5(6):729–736

    Article  PubMed  PubMed Central  Google Scholar 

  19. Pfyffer GE (1992) Serratia fonticola as an infectious agent. Eur J Clin Microbiol Infect Dis 11(2):199–200

    Article  CAS  PubMed  Google Scholar 

  20. Soler C, Samson T, Hernandez E, Hervé V, De Revel T (2000) Serratia fonticola, germe opportuniste: à propos d’une observation chez un immunodéprimé. Med Maladies Infect 30(9):599–600

    Article  Google Scholar 

  21. Stock I, Burak S, Sherwood KJ, Grüger T, Wiedemann B (2003) Natural antimicrobial susceptibilities of strains of ‘unusual’ Serratia species: S. ficaria, S. fonticola, S. odorifera, S. plymuthica and S. rubidaea. J Antimicrob Chemoth 51(4):865–885

    Article  CAS  Google Scholar 

  22. Tasic S, Obradovic D, Tasic I (2013) Characterization of Serratia fonticola, an opportunistic pathogen isolated from drinking water. Arch Biol Sci 65(3):899–904

    Article  Google Scholar 

  23. Wang JC (1985) DNA topoisomerases. Annu Rev Biochem 2(54):665–697

    Article  Google Scholar 

  24. Yamamoto S, Harayama S (1996) Phylogenetic analysis of acinetobacter strains based on the nucleotide sequences of gyrB Genes and on the amino acid sequences of their products. Int J Syst Bacteriol 46(2):506–511

    Article  CAS  PubMed  Google Scholar 

  25. Zhu H, Sun SJ, Dang HY (2008) PCR detection of Serratia spp. using primers targeting pfs and luxS genes involved in AI-2-dependent quorum sensing. Curr Microbiol 57(4):326–330

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the 13th Five-Year State Key Development Program [Grant Number 2016YFD0501310], the National Natural Science Foundation of China [Grant Number 31272606], Science Program of General Administration of Quality Supervision, Inspection and Quarantine of P.R.C [Program Number 2016IK030] and Science Program of Fujian Entry-Exit Inspection and Quarantine Bureau of P.R.C [Program Number FJ2015-JS002].

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Author notes

  1. Jing Hua Ruan and Wu Jun Wang have contributed equally to this work.

Authors and Affiliations

  1. Fujian Key Laboratory of Traditional Chinese Veterinary Medicine and Animal Health, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China

    Jing Hua Ruan, Wu Jun Wang, Li Yun Wu, Yi Fan Huang & Dao Jin Yu

  2. College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, Fujian, China

    Wu Jun Wang

  3. Fujian Key Laboratory for Technology Research of Inspection and Quarantine, Fujian Entry-Exit Inspection and Quarantine Bureau, Fuzhou, 350001, Fujian, China

    Wu Jun Wang, Ti Yin Zhang, Quan Yang Bai, Teng Zheng & Zhi Deng Zhang

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  1. Jing Hua Ruan
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  2. Wu Jun Wang
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Correspondence to Yi Fan Huang or Dao Jin Yu.

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Ruan, J.H., Wang, W.J., Zhang, T.Y. et al. Rapid Detection of Serratia fonticola by TaqMan Quantitative Real-Time PCR Using Primers Targeting the gyrB Gene. Curr Microbiol 74, 1343–1348 (2017). https://doi.org/10.1007/s00284-017-1323-x

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  • DOI: https://doi.org/10.1007/s00284-017-1323-x

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