Abstract
The control and prevention of meticillin-resistant Staphylococcus aureus (MRSA) is a major challenge for healthcare establishments, especially as this pathogen continues to evolve. The emergence and spread of community associated MRSA producing Panton–Valentine leukocidin (PVL) causing severe, sometimes fatal, infections in otherwise healthy people is a significant cause of concern. Patient screening to detect MRSA is now widely used as part of an effective control program to limit the spread of this pathogen. Real-time PCR targeting specific MRSA markers offers a rapid alternative to conventional methods enabling earlier intervention, such as patient isolation and decolonization treatment. Herein we describe a multiplex real-time assay that combines primers and probes to detect MRSA and the genes for PVL to provide a rapid and informative assay.
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References
Vandenesch F, Naimi T, Enright MC, Lina G, Nimmo GR, Heffernan H, Liassine N, Bes M, Greenland T, Reverdy ME, Etienne J (2003) Community-acquired methicillin-resistant Staphylococcus aureus carrying Panton-Valentine leukocidin genes: worldwide emergence. Emerg Infect Dis 9:978–984
Hardy K, Price C, Szczepura A, Gossain S, Davies R, Stallard N, Shabir S, McMurray C, Bradbury A, Hawkey PM (2010) Reduction in the rate of methicillin-resistant Staphylococcus aureus acquisition in surgical wards by rapid screening for colonisation: a prospective, cross-over study. Clin Microbiol Infect 16(4):333–339
Jog S, Cunningham R, Cooper S, Wallis M, Marchbank A, Vasco-Knight P, Jenks PJ (2008) Impact of preoperative screening for methicillin-resistant Staphylococcus aureus by real-time polymerase chain reaction in patients undergoing cardiac surgery. J Hosp Infect 69:124–130
Keshtgar MR, Khalilli A, Coen PG, Carder C, Macrae B, Jeanes A, Folan P, Baker D, Wren M, Wilson AP (2008) Impact of rapid molecular screening for methicillin-resistant Staphylococcus aureus in surgical wards. Br J Surg 95:381–386
Huletsky A, Giroux R, Rossbach V, Gagnon M, Vaillancourt M, Bernier M, Gagnon F, Truchon K, Bastien M, Picard FJ, van Belkum A, Ouelle M, Roy PH, Bergeron MG (2004) New real-time PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus directly from specimens containing a mixture of staphylococci. J Clin Microbiol 42:1875–1884
Prevost G, Cribier B, Couppie P, Petiau P, Supersac G, Finck-Barbancon V, Monteil H, Piemont Y (1995) Panton-Valentine leucocidin and gamma-hemolysin from Staphylococcus aureus ATCC 49775 are encoded by distinct genetic loci and have different biological activities. Infect Immun 63:4121–4129
Prevost GL, Mourey L, Colin DA, Menestrina G (2001) Staphylococcal pore-forming toxins. Curr Top Microbiol Immunol 257:53–83
Ellington MJ, Perry C, Ganner M, Warner M, McCormick Smith I, Hill RL, Shallcross L, Khatri K, Sabersheikh S, Holmes A, Cookson BD, Kearns AM (2009) Clinical and molecular epidemiology of ciprofloxacin-susceptible MRSA encoding PVL in England and Wales. Eur J Clin Microbiol Infect Dis 28:1113–1121
Seybold U, Kourbatova EV, Johnson JG, Halvosa SJ, Wang YF, King MD, Ray SM, Blumberg HM (2006) Emergence of community-associated methicillin-resistant Staphylococcus aureus USA 300 genotype as a major cause of health care—associated blood stream infections. Clin Infect Dis 42:647–666
Gonzalez BE, Rueda AM, Shelburne SA, Musher DM, Hamill RJ, Hulten KG (2006) Community-associated strains of methicillin-resistant Staphylococcus aureus as the cause of healthcare-associated infection. Infect Control Hosp Epidemiol 27:1051–1056
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 Staphylococus aureus. J Clin Microbiol 43:6147–6149
Renwick L, Hardie A, Girvan EK, Smith M, Leadbetter G, Claas E, Morrison D, Gibb AP, Dave J, Templeton KE (2008) Detection of meticillin-resistant Staphylococcus aureus and Panton-Valentine leukocidin directly from clinical samples and the development of a multiplex assay using real-time polymerase chain reaction. Eur J Clin Microbiol Infect Dis 27:791–796
Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J (1990) Rapid and simple method for extraction of nucleic acids. J Clin Microbiol 28:495–503
Shore AC, Rossney AS, O’Connell B, Herra CM, Sullivan DJ, Humphreys H, Coleman DC (2008) Detection of staphylococcal cassette chromosome mec-associated DNA elements in multiresistant methicillin-susceptible Staphylococcus aureus (MSSA) and identification of Staphylococcus epidermidis ccrAB4 in both methcillin-resistant S. aureus and MSSA. Antimicrob Agents Chemother 52:4407–4419
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Renwick, L., Holmes, A., Templeton, K. (2013). Multiplex Real-Time PCR Assay for the Detection of Meticillin-Resistant Staphylococcus aureus and Panton–Valentine Leukocidin from Clinical Samples. In: Wilks, M. (eds) PCR Detection of Microbial Pathogens. Methods in Molecular Biology, vol 943. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-353-4_6
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DOI: https://doi.org/10.1007/978-1-60327-353-4_6
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Publisher Name: Humana Press, Totowa, NJ
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