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Volume 12, Number 2—February 2006
Dispatch

Sequencing and Staphylococci Identification

Alexander Mellmann*Comments to Author , Karsten Becker*, Christof von Eiff*, Ursula Keckevoet*, Peter Schumann†, and Dag Harmsen*
Author affiliations: *University Hospital Münster, Münster, Germany; †Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany

Main Article

Table 1

Primers used for amplification and partial sequencing of the partial staphylococcal RNA polymerase B (rpoB) gene

Primer Application Primer sequence (5´→3´) Annealing temperature (°C) Reference
Staph rpoB 1418f* Amplification and sequencing CAA TTC ATG GAC CAA GC 52 Modified from 7
Staph rpoB 3554r Amplification CCG TCC CAA GTC ATG AAA C 52 7
Staph rpoB 1975r* Sequencing GCI ACI TGI TCC ATA CCT GT 52 Modified from 7
Staph rpoB 1876r*† Sequencing GAG TCA TCI TTY TCT AAG AAT GG 52 This study

*Primers are numbered from the 3´ end of the primer on the forward strand of Staphylococcus aureus (GenBank accession no. X64172).
†Primer was used for sequencing when primer Staph rpoB 1975r did not work.

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References
  1. von Eiff  C, Peters  G, Heilmann  C. Pathogenesis of infections due to coagulase-negative staphylococci. Lancet Infect Dis. 2002;2:67785. DOIPubMedGoogle Scholar
  2. Seifert  H, Wisplinghoff  H, Schnabel  P, von Eiff  C. Small colony variants of Staphylococcus aureus and pacemaker-related infection. Emerg Infect Dis. 2003;9:13168.PubMedGoogle Scholar
  3. Becker  K, Harmsen  D, Mellmann  A, Meier  C, Schumann  P, Peters  G, Development and evaluation of a quality-controlled ribosomal sequence database for 16S ribosomal DNA-based identification of Staphylococcus species. J Clin Microbiol. 2004;42:498895. DOIPubMedGoogle Scholar
  4. Kwok  AY, Su  SC, Reynolds  RP, Bay  SJ, Av-Gay  Y, Dovichi  NJ, Species identification and phylogenetic relationships based on partial HSP60 gene sequences within the genus Staphylococcus. Int J Syst Bacteriol. 1999;49:118192. DOIPubMedGoogle Scholar
  5. Poyart  C, Quesne  G, Boumaila  C, Trieu-Cuot  P. Rapid and accurate species-level identification of coagulase-negative staphylococci by using the sodA gene as a target. J Clin Microbiol. 2001;39:4296301. DOIPubMedGoogle Scholar
  6. Drancourt  M, Raoult  D. rpoB gene sequence-based identification of Staphylococcus species. J Clin Microbiol. 2002;40:13338. DOIPubMedGoogle Scholar
  7. Euzeby  JP. List of bacterial names with standing in nomenclature: a folder available on the internet. Int J Syst Bacteriol. 1997;47:5902. DOIPubMedGoogle Scholar
  8. Trülzsch  K, Rinder  H, Trcek  J, Bader  L, Wilhelm  U, Heesemann  J. 'Staphylococcus pettenkoferi' a novel staphylococcal species isolated from clinical specimens. Diagn Microbiol Infect Dis. 2002;43:17582. DOIPubMedGoogle Scholar
  9. Harmsen  D, Karch  H. 16S rDNA for diagnosing pathogens: a living tree. ASM News. 2004;70:1924.
  10. Clarridge  JE. Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev. 2004;17:84062. DOIPubMedGoogle Scholar
  11. Drancourt  M, Roux  V, Fournier  P, Raoult  D. rpoB gene sequence-based identification of aerobic gram-positive cocci of the genera Streptococcus, Enterococcus, Gemella, Abiotrophia, and Granulicatella. J Clin Microbiol. 2004;42:497504. DOIPubMedGoogle Scholar
  12. Mollet  C, Drancourt  M, Raoult  D. rpoB sequence analysis as a novel basis for bacterial identification. Mol Microbiol. 1997;26:100511. DOIPubMedGoogle Scholar
  13. Khamis  A, Raoult  D, La Scola  B. Comparison between rpoB and 16S rRNA gene sequencing for molecular identification of 168 clinical isolates of Corynebacterium. J Clin Microbiol. 2005;43:19346. DOIPubMedGoogle Scholar
  14. Blackwood  KS, Turenne  CY, Harmsen  D, Kabani  AM. Reassessment of sequence-based targets for identification of Bacillus species. J Clin Microbiol. 2004;42:162630. DOIPubMedGoogle Scholar
  15. Cook  VJ, Turenne  CY, Wolfe  J, Pauls  R, Kabani  A. Conventional methods versus 16S ribosomal DNA sequencing for identification of nontuberculous mycobacteria: cost analysis. J Clin Microbiol. 2003;41:10105. DOIPubMedGoogle Scholar

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