Rapid detection of Panton–Valentine leukocidin-positive Staphylococcus aureus by real-time PCR targeting the lukS-PV gene

  • U. ReischlEmail author
  • M. J. Tuohy
  • G. S. Hall
  • G. W. Procop
  • N. Lehn
  • H. Linde
Concise Article


In order to assess the speed and accuracy of a real-time PCR assay targeting the lukS-PV gene of Panton–Valentine leukocidin (PVL)-positive Staphylococcus aureus, 700 S. aureus strains were tested and the results were compared to those achieved with block cycler PCR. Cross-reactivity was tested with 166 other bacterial species. Using this homogeneous real-time PCR assay format, the presence or absence of genetic information for PVL, which is also found in community-associated methicillin-resistant S. aureus, was correctly identified from pure culture and directly in various types of clinical specimens.


Hybridization Probe Necrotizing Fasciitis SCCmec Type Wound Swab MLST Type 
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  1. 1.
    Baba T, Takeuchi F, Kuroda M, Yuzawa H, Aoki K, Oguchi A, Nagai Y, Iwama N, Asano K, Naimi T, Kuroda H, Cui L, Yamamoto K, Hiramatsu K (2002) Genome and virulence determinants of high virulence community-acquired MRSA. Lancet 359:1819–1827PubMedCrossRefGoogle Scholar
  2. 2.
    Pragman AA, Schlievert PM (2004) Virulence regulation in Staphylococcus aureus: the need for in vivo analysis of virulence factor regulation. FEMS Immunol Med Microbiol 42:147–154PubMedCrossRefGoogle Scholar
  3. 3.
    von Eiff C, Friedrich AW, Peters G, Becker K (2004) Prevalence of genes encoding for members of the staphylococcal leukotoxin family among clinical isolates of Staphylococcus aureus. Diagn Microbiol Infect Dis 49:157–162CrossRefGoogle Scholar
  4. 4.
    Lina G, Piemont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V, Vandenesch F, Etienne J (1999) Involvement of Panton–Valentine leukocidin-producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis 29:1128–1132PubMedCrossRefGoogle Scholar
  5. 5.
    Diep BA, Sensabaugh GF, Somboona NS, Carleton HA, Perdreau-Remington F (2004) Widespread skin and soft-tissue infections due to two methicillin-resistant Staphylococcus aureus strains harboring the genes for Panton–Valentine leucocidin. J Clin Microbiol 42:2080–2084PubMedCrossRefGoogle Scholar
  6. 6.
    Martinez-Aguilar G, Avalos-Mishaan A, Hulten K, Hammerman W, Mason EO Jr, Kaplan SL (2004) Community-acquired, methicillin-resistant and methicillin-susceptible Staphylococcus aureus musculoskeletal infections in children. Pediatr Infect Dis J 23:701–706PubMedCrossRefGoogle Scholar
  7. 7.
    Ribeiro A, Dias C, Silva-Carvalho MC, Berquo L, Ferreira FA, Santos RN, Ferreira-Carvalho BT, Figueiredo AM (2005) First report of infection with community-acquired methicillin-resistant Staphylococcus aureus in South America. J Clin Microbiol 43:1985–1988PubMedCrossRefGoogle Scholar
  8. 8.
    Gillet Y, Issartel B, Vanhems P, Fournet JC, Lina G, Bes M, Vandenesch F, Piemont Y, Brousse N, Floret D, Etienne J (2002) Association between Staphylococcus aureus strains carrying gene for Panton–Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients. Lancet 359:753–759PubMedCrossRefGoogle Scholar
  9. 9.
    Hsu L, Koh T, Anantham D, Kurup A, Ping Wah Chan K, Tan B (2004) Panton–Valentine leukocidin-positive Staphylococcus aureus, Singapore. Emerg Infect Dis 10:1509–1510PubMedGoogle Scholar
  10. 10.
    Miller LG, Perdreau-Remington F, Rieg G, Mehdi S, Perlroth J, Bayer AS, Tang AW, Phung TO, Spellberg B (2005) Necrotizing fasciitis caused by community-associated methicillin-resistant Staphylococcus aureus in Los Angeles. N Engl J Med 352:1445–1453PubMedCrossRefGoogle Scholar
  11. 11.
    Panton PN, Valentine FCO (1932) Staphylococcal toxin. Lancet 222:506–508CrossRefGoogle Scholar
  12. 12.
    Kaneko J, Kamio Y (2004) Bacterial two-component and hetero-heptameric pore-forming cytolytic toxins: structures, pore-forming mechanism, and organization of the genes. Biosci Biotechnol Biochem 68:981–1003PubMedCrossRefGoogle Scholar
  13. 13.
    Genestier AL, Michallet MC, Prevost G, Bellot G, Chalabreysse L, Peyrol S, Thivolet F, Etienne J, Lina G, Vallette FM, Vandenesch F, Genestier L (2005) Staphylococcus aureus Panton–Valentine leukocidin directly targets mitochondria and induces Bax-independent apoptosis of human neutrophils. J Clin Invest 115:3117–3127PubMedCrossRefGoogle Scholar
  14. 14.
    Prevost G, Couppie P, Prevost P, Gayet S, Petiau P, Cribier B, Monteil H, Piemont Y (1995) Epidemiological data on Staphylococcus aureus strains producing synergohymenotropic toxins. J Med Microbiol 42:237–245PubMedCrossRefGoogle Scholar
  15. 15.
    Melles DC, Gorkink RF, Boelens HA, Snijders SV, Peeters JK, Moorhouse MJ, van der Spek PJ, van Leeuwen WB, Simons G, Verbrugh HA, van Belkum A (2004) Natural population dynamics and expansion of pathogenic clones of Staphylococcus aureus. J Clin Invest 114:1732–1740PubMedCrossRefGoogle Scholar
  16. 16.
    Okuma K, Iwakawa K, Turnidge JD, Grubb WB, Bell JM, O’Brien FG, Coombs GW, Pearman JW, Tenover FC, Kapi M, Tiensasitorn C, Ito T, Hiramatsu K (2002) Dissemination of new methicillin-resistant Staphylococcus aureus clones in the community. J Clin Microbiol 40:4289–4294PubMedCrossRefGoogle Scholar
  17. 17.
    Holmes A, Ganner M, McGuane S, Pitt TL, Cookson BD, Kearns AM (2005) Staphylococcus aureus isolates carrying Panton–Valentine leucocidin genes in England and Wales: frequency, characterization, and association with clinical disease. J Clin Microbiol 43:2384–2390PubMedCrossRefGoogle Scholar
  18. 18.
    Linde H, Wagenlehner F, Strommenger B, Drubel I, Tanzer J, Reischl U, Raab U, Holler C, Naber KG, Witte W, Hanses F, Salzberger B, Lehn N (2005) Healthcare-associated outbreaks and community-acquired infections due to MRSA carrying the Panton–Valentine leucocidin gene in southeastern Germany. Eur J Clin Microbiol Infect Dis 24:419–422PubMedCrossRefGoogle Scholar
  19. 19.
    Centers for Disease Control (1999) Four pediatric deaths from community-acquired methicillin-resistant Staphylococcus aureus: Minnesota and North Dakota, 1997–1999. MMWR 48:707–710Google Scholar
  20. 20.
    Charlebois ED, Bangsberg DR, Moss NJ, Moore MR, Moss AR, Chambers HF, Perdreau-Remington F (2002) Population-based community prevalence of methicillin-resistant Staphylococcus aureus in the urban poor of San Francisco. Clin Infect Dis 34:425–433PubMedCrossRefGoogle Scholar
  21. 21.
    Fridkin SK, Hageman JC, Morrison M, Sanza LT, Como-Sabetti K, Jernigan JA, Harriman K, Harrison LH, Lynfield R, Farley MM (2005) Methicillin-resistant Staphylococcus aureus disease in three communities. N Engl J Med 352:1436–1444PubMedCrossRefGoogle Scholar
  22. 22.
    Witte W, Braulke C, Cuny C, Strommenger B, Werner G, Heuck D, Jappe U, Wendt C, Linde HJ, Harmsen D (2004) Emergence of methicillin-resistant Staphylococcus aureus with Panton–Valentine leukocidin genes in central Europe. Eur J Clin Microbiol Infect Dis 24:1–5CrossRefGoogle Scholar
  23. 23.
    Deurenberg RH, Vink C, Driessen C, Bes M, London N, Etienne J, Stobberingh EE (2004) Rapid detection of Panton–Valentine leukocidin from clinical isolates of Staphylococcus aureus strains by real-time PCR. FEMS Microbiol Lett 240:225–228, Nov. 15PubMedCrossRefGoogle Scholar
  24. 24.
    Johnsson D, Molling P, Stralin K, Soderquist B (2004) Detection of Panton–Valentine leukocidin gene in Staphylococcus aureus by LightCycler PCR: clinical and epidemiological aspects. Clin Microbiol Infect 10:884–889PubMedCrossRefGoogle Scholar
  25. 25.
    Reischl U, Youssef MT, Kilwinski J, Lehn N, Zhang WL, Karch H, Strockbine NA (2002) Real-time fluorescence PCR assays for detection and characterization of Shiga toxin, intimin, and enterohemolysin genes from Shiga toxin-producing Escherichia coli. J Clin Microbiol 40:2555–2565PubMedCrossRefGoogle Scholar
  26. 26.
    Reischl U, Linde HJ, Metz M, Leppmeier B, Lehn N (2000) Rapid identification of methicillin-resistant Staphylococcus aureus and simultaneous species confirmation using real-time fluorescence PCR. J Clin Microbiol 38:2429–2433PubMedGoogle Scholar
  27. 27.
    Robinson DA, Enright MC (2004) Multilocus sequence typing and the evolution of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect 10:92–97PubMedCrossRefGoogle Scholar
  28. 28.
    Obed A, Schnitzbauer AA, Bein T, Lehn N, Linde HJ, Schlitt HJ (2006) Fatal pneumonia caused by Panton–Valentine leucocidine-positive methicillin-resistant Staphylococcus aureus (PVL-MRSA) transmitted from a healthy donor in living-donor liver transplantation. Transplantation 81:121–124PubMedCrossRefGoogle Scholar
  29. 29.
    McDonald RR, Antonishyn NA, Hansen T, Snook LA, Nagle E, Mulvey MR, Levett PN, Horsman GB (2005) Development of a triplex real-time PCR assay for detection of Panton–Valentine leukocidin toxin genes in clinical isolates of methicillin-resistant Staphylococcus aureus. J Clin Microbiol 43:6147–6149PubMedCrossRefGoogle Scholar
  30. 30.
    Nakagawa S, Taneike I, Mimura D, Iwakura N, Nakayama T, Emura T, Kitatsuji M, Fujimoto A, Yamamoto T (2005) Gene sequences and specific detection for Panton–Valentine leukocidin. Biochem Biophys Res Commun 328:995–1002PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • U. Reischl
    • 1
    Email author
  • M. J. Tuohy
    • 2
  • G. S. Hall
    • 2
  • G. W. Procop
    • 2
  • N. Lehn
    • 1
  • H. Linde
    • 1
  1. 1.Institute of Medical Microbiology and HygieneUniversity Hospital of RegensburgRegensburgGermany
  2. 2.Clinical MicrobiologyCleveland Clinic FoundationClevelandUSA

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