Abstract
Staphylococcus lugdunensis is an unusually virulent coagulase‐negative species, associated with severe infections. The present report describes the development of a single‐step, species‐specific PCR protocol for S. lugdunensis identification. fbl gene, encoding a fibrinogen‐binding adhesin, was exploited and assessed as a suitable nucleic acid target. The gene was detected in all 17 S. lugdunensis isolates examined, while no amplification product was obtained from 98 isolates representing 11 staphylococcal and 17 nonstaphylococcal species. Forty‐seven percent of the S. lugdunensis strains produced a positive slide coagulase reaction, which is consistent with varying levels of Fbl protein expression within the species. J. Clin. Lab. Anal. 24:119–122, 2010. © 2010 Wiley‐Liss, Inc.
Keywords: fbl gene, identification, PCR, Staphylococcus lugdunensis, fbl protein, expression
REFERENCES
- 1. Anguera I, Del Rio A, Miro JM, et al. Staphylococcus lugdunensis infective endocarditis: Description of 10 cases and analysis of native valve, prosthetic valve, and pacemaker lead endocarditis clinical profiles. Heart 2005;91:e10. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2. Frank KL, Del Pozo JL, Patel R. From clinical microbiology to infection pathogenesis: how daring to be different works for Staphylococcus lugdunensis . Clin Microbiol Rev 2008;21:111–133. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. Tan YT, Ng SY, He J. Microbiological characteristics, presumptive identification and antibiotic susceptibilities of Staphylococcus lugdunensis . J Clin Microbiol 2008;46:2393–2395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. Mateo M, Maestre JR, Aguilar L, et al. Genotypic versus phenotypic characterization, with respect to susceptibility and identification, of 17 clinical isolates of Staphylococcus lugdunensis . J Antimicrob Chemother 2005;56:287–291. [DOI] [PubMed] [Google Scholar]
- 5. Noguchi N, Goto K, Ro T, et al. Using the tannase gene to rapidly and simply identify Staphylococcus lugdunensis . Diagn Microbiol Infect Dis 2009. [Epub ahead of print]. [DOI] [PubMed] [Google Scholar]
- 6. Mitchell J, Tristan A, Foster TJ. Characterization of the fibrinogen‐binding surface protein Fbl of Staphylococcus lugdunensis. Microbiology 2004;150:3831–3841. [DOI] [PubMed] [Google Scholar]
- 7. Nilsson M, Bjerketorp J, Guss B, Frykberg L. A fibrinogen‐binding protein of Staphylococcus lugdunensis . FEMS Microbiol Lett 2004;241:87–93. [DOI] [PubMed] [Google Scholar]
- 8. Hiraishi A. Direct automated sequencing of 16S rDNA amplified by polymerase chain reaction from bacterial cultures without DNA purification. Lett Appl Microbiol 1992;15:210–213. [DOI] [PubMed] [Google Scholar]
- 9. Kontos F, Petinaki E, Spiliopoulou I, Maniati M, Maniatis AN. Evaluation of a novel method based on PCR restriction fragment length polymorphism analysis of the tuf gene for the identification of Staphylococcus species. J Microbiol Methods 2003;55:465–469. [DOI] [PubMed] [Google Scholar]
- 10. Casas I, Powell L, Klapper PE, Cleator GM. New method for the extraction of viral RNA and DNA from cerebrospinal fluid for use in the polymerase chain reaction assay. J Virol Methods 1995;53:25–26. [DOI] [PubMed] [Google Scholar]
- 11. Ghebremedhin B, Layer F, König W, König B. Genetic classification and distinguishing of Staphylococcus species based on different partial gap, 16S rRNA, hsp60, rpoB, sodA, and tuf gene sequences. J Clin Microbiol 2008;46:1019–1025. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12. Barros EM, Iório NL, Bastos MdoC, dos Santos KR, Giambiagi‐deMarval M. Species‐level identification of clinical staphylococcal isolates based on polymerase chain reaction–restriction fragment length polymorphism analysis of a partial groEL gene sequence. Diagn Microbiol Infect Dis 2007;59:251–257. [DOI] [PubMed] [Google Scholar]
- 13. Couto I, Pereira S, Miragaia M, Sanches IS, de Lencastre H. Identification of clinical staphylococcal isolates from humans by internal transcribed spacer PCR. J Clin Microbiol 2001;39:3099–3103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14. Andollina A, De Cesare A, Bertoni G, Modelli L, Manfreda G. Identification and genetic characterisation of orthopaedic Staphylococcus isolates collected in Italy by automated EcoRI ribotyping. FEMS Microbiol Lett 2004;234:275–280. [DOI] [PubMed] [Google Scholar]
- 15. Skow A, Mangold KA, Tajuddin M, et al. Species‐level identification of staphylococcal isolates by real‐time PCR and melt curve analysis. J Clin Microbiol 2005;43:2876–2880. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. Giammarinaro P, Leroy S, Chacornac JP, Delmas J, Talon R. Development of a new oligonucleotide array to identify staphylococcal strains at species level. J Clin Microbiol 2005;43:3673–3680. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. Personne P, Bes M, Lina G, Vandenesch F, Brun Y, Etienne J. Comparative performances of six agglutination kits assessed by using typical and atypical strains of Staphylococcus aureus . J Clin Microbiol 1997;35:1138–1140. [DOI] [PMC free article] [PubMed] [Google Scholar]