DOI: | Full Text: [PDF] [HTML]

Genotypic and phenotypic characteristics of Methicillin-resistant Staphylococcus aureus (MRSA) strains, isolated on three different geography locations

Maja Ostojić, Mirsada Hukić


Staphylococcus aureus is a major cause of hospital-acquired infections worldwide. Increased frequency of methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients and possibility of vancomycin resistance requires rapid and reliable characterization of isolates and control of MRSA spread in hospitals. Typing of isolates helps to understand the route of a hospital pathogen spread. The aim of this study was to investigate and compare genotypic and phenotypic characteristics of MRSA samples on three different geography locations. In addition, our aim was to evaluate three different methods of MRSA typing: spa-typing, agr-typing and GenoType MRSA.  We included 104 samples of MRSA, isolated in 3 different geographical locations in clinical hospitals in Zagreb, Mostar, and Heidelberg, during the period of six months. Genotyping and phenotyping were done by spa-typing, agr-typing and dipstick assay GenoType MRSA. We failed to type all our samples by spa-typing.  The most common spa-type in clinical hospital Zagreb was t041, in Mostar t001, and in Heidelberg t003.We analyzed 102/104 of our samples by agr-typing method. We did not find any agr-type IV in our locations. We analyzed all our samples by the dipstick assay GenoType MRSA. All isolates in our study were MRSA strains. In Zagreb there were no positive strains to PVL gene. In Mostar we have found 5/25 positive strains to PVL gene, in Heidelberg there was 1/49. PVL positive isolates were associated with spa-type t008 and agr-type I, thus, genetically, they were community-associated MRSA (CA-MRSA). Dipstick assay GenoType MRSA has demonstrated sufficient specificity, sensibility, simple performance and low cost, so we could introduce it to work in smaller laboratories. Using this method may expedite MRSA screening, thus preventing its spread in hospitals.


Methicillin resistant Staphylococcus aureus; spa-typing; agr-typing; GenoType MRSA; infection control; MRSA

Full Text:



Waldvogel FA. Staphylococcus aureus (including toxic shock syndrome) In: Mandell GL, Bennett JE, Dolin R eds. Principles and Practice of Infectious Diseases, 5th ed. London, Churchill Livingstone 2000; 2069-2089

Gram-positive cocci In: KonemanEW, Allen SD, Janda WM, Schreckenberger PC, Winn WC eds Color Atlas and Textbook of Diagnostic Microbiology 6th ed. Philadelphia: J.B. Lippincot Co., 2006; 623-662.

Staphylococcus and related organisms In: Murray PR, Rosenthal KS, Pfaller MA. Medical Microbiology 5th ed. Philadelphia (Pe): Elsevier Mosby, 2005; 221-236.

Ostojić M. Staphylococcus, Micrococcus and the other catalasa-positive cocci In: Uzunović-Kamberović S, eds Medical microbiology, Zenica, Press Fojnica, 2009; 259-267.

Hukić M. Gram-positive cocci. In: Hukić M et al. Bacteriology. Sarajevo: Jež d.o.o., 2005; 163-166.

Reverdy ME1, Jarraud S, Bobin-Dubreux S, Burel E, Girardo P, Lina G, et al. Incidence of Staphylococcus aureus with reduced susceptibility to glycopeptides in two French hospitals. ClinMicrobiol Infect 2001; 7(5): 267-272.

Bradley SF. Methicillin-resistant Staphylococcus aureus infection. Infect Dis 1992; 8:853-868.

Boyce J. Methicillin-resistant Staphylococcus aureus infections: a growing control problem. Infect Control 1980; 1:335-336.

French GL. A short history of methicillin-resistant Staphylococcus aureus. Abstracts book: 2nd Croatian Symposium on Bacterial Resistance to Antibiotics. Croatian Academy of medical sciences, Zagreb; 1997:pp13-17.

Katayama Y, Ito T, Hiramatsu K. A new class of genetic element, staphylococcus cassette chromosome mec, encodes methicillin resistance in Staphylococcus aureus. Antimicrob Agents Chemother 2000; 44: 1549–1555.

Pantosti A, Venditti M. What is MRSA? EurRespir J 2009;34:1190–1196.

Ostojić M, Glibić I, Parčina M. Epidemiological typing of Methicillin-resistant Staphylococcus aureus (MRSA) by PFGE. Acta Medica Saliniana 2005; 34 (Supl 1): 1-98.

European Antimicrobial Resistance Surveillance System. EARSS Annual Report 2007. Available from: Accessed 14th January 2015.

Tiemersma EW, Bronzwaer SL, Lyytikainen O, Degener JE, Schrijnemakers P, Bruinsma N, et al. Methicillin-resistant Staphylococcus aureus in Europe, 1999–2002. Emerg Infect Dis 2004; 10: 1627–1634.

Budimir A. Detection and typing methods of methicillin-resistant Staphylococcus aureus strains. Medical Sciences 37(2012): 73-88.

Sievert DM, Rudrik JT, Patel JB, McDonald LC, Wilkins MJ, Hageman JC. Vancomycin-resistant Staphylococcus aureus in the United States, 2002–2006. Clin Infect Dis 2008;46:668–674.

IFIC, International Federation of Infection Control; Referral center for infections control of Republic Croatia. Infection control: Basic concept ant training. Merkur A.B.D. Zagreb, Croatia, 2004 , pp 40-51.

Ostojić M. Methicillin resistant Staphylococcus aureus (MRSA). In: Hukić M (ed). Prevention and control of infections in healthcare facilities, 1st ed. Sarajevo: TDP Sarajevo; 2010, pp. 291-295.

Bischoff WE and Edmond MB. Staphylococcus aureus. In: Wenzel PR (ed). A guide to infection control in the Hospital. AGM d.o.o. B.C. Decker Inc Hamilton-London 1998; 173-176

Damani N, Keyes J, Campbell H: Infection Control Manual, Greenwich Media Ltd, UK 2004; 6.1-6.7.)

Miljković-Selimović B, Kocić B, Babić T, RistićLj. Bacterial typing methods, Acta Fac Med Naiss 2009; 26: 225-233.

De Sousa MA, De Lencastre H. Bridges from hospitals to the laboratory: genetic portraits od methicillin-resistant Staphylococcus aureus clones. FEMS Immunol Med Microbiol 40, 101 (2004).

DeurenbergRH, Vink C, Kalenić S, Friedrich AW, Bruggeman CA and Stobberingh EE. The molecular evolution of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect 2007;13: 222–235.

Stranden A, Frei R, Widmer AF. Molecular typing of methicillin-resistant Staphylococcus aureus; can PCR replace pulsed-field gel electrophoresis? J Clin Microbiol 2003: 41: 3181-6

Frenay HM, Bunschoten AE, Schouls LM, Van Leeuwen WJ, Vanderbroucke-Grauls CM, Verhoef J, et al. Molecular typing of methicillin-resistant Staphylococcus aureus on the basis of protein A gene polymorphism. Eur J ClinMicrobiol Infect Dis 1996; 15: 60–64.

Stromenger B, Kettlitz C, Weniger T, Harmsen D, Friedrich A, Witte W. Assignment of Staphylococcus isolates to groups by spa typing, SmaImacrorestriction analysis, and multilocus sequence typing. J ClinMicrobiol. 2006; 44:2533-40.

Harmsen D, Claus H, Witte W, Rothgänger J, Turnwald D, Vogel U et al. Typing of methicillin-resistant Staphylococcus aureus in a university hospital setting by using novel software for spa repeat determination and database management. J. Clin. Microbiol. 2003; 41:5442-5448.

Otte KM, Jenner S, Wulffen HV. Identification of methicillin-resistant Staphylococcus aureus (MRSA): Comparison of a new molecular genetic test kit (GenoType MRSA) with standard diagnostic methods. Clin Lab 2005; 51(7-8): 389-39.

Mc Dougal LK, Steward CD, Killgore GE, Chaitram JM, Mc Allister SK, Tenover FC . Pulsed-field gel electrophoresis typing of oxacillin-resistant Staphylococcus aureus isolates from the United States: Establishing a national database. J Clin Microbiol 2003; 41: 5113-5120.

Ostojić M. Epidemiologic genotyping of methicillin-resistant Staphylococcus aureus (MRSA) by pulsed-field gel electrophoresis (PFGE). Bosn J Basic Med Sci 2008; 8(3):259-265

Vindel A, Cuevas O, Cercenado E, Marcos C, Bautista V, Castellares C, et al. Methicillin-Resistant Staphylococcus aureus in Spain: molecular epidemiology and utility of different typing methods. J Clin Microbiol. 2009; 47(6): 1620-1627.

Jarraud S, Mougel C, Thioulouse J, Lina G, Meugnier H, Forey F et al. Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups (alleles), and human disease. Infect Immun 2002; 70(2): 631-341.

Manago K, Nishi J, Wakimoto N, Miyanohara H, Jay Sarantuya MT, Tokuda K et al. Biofilm formation by and accessory gene regulator typing of methicillin-resistant Staphylococcus aureus strains recovered from patients with nosocomial infections. Infect Control Hosp Epidemiol. 2006;27:188–190.

Kearns AM, Seiders PR, Weeler J, Freeman E, and Steward M. Rapid detection of methicillin-resistant staphylococci by multiplex PCR. J. Hospital. Infection 1999; 43: 33-37.

Deurenberg RH, Vink C, Kalenić S, Friedrich AW, Bruggeman CA and Stobberingh EE.: The molecular evolution of methicillin-resistant Staphylococcus aureus. ClinMicrobiolInfect 2007, 13: 222–235.

David MZ, Daum RS. Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic. ClinMicrobiol Rev 2010;23:616-87.

Kalenić S, Payerl Pal M, Vlahović Palčevski V, Horvatić J, Meštrović T, Baršić B et al. Guidelines for preventing, control and treatment of methicillin-resistant Staphylococcus aureus (MRSA) associated infections. Electronic form only 2011. Available from: Accessed 20 January 2015.

Witte W, Strommenger B, Cuny C, Heuck D, Nuebel U. Methicillin-resistant Staphylococcus aureus containing the Panton-Valentine leucocidin gene in Germany in 2005 and 2006. J AntimicrobChemother 2007;60:1258-63.

Blanco R, Tristan A, Ezpeleta G, Rhod Larsen A, Bes M, Etienne J et al. Molecular Epidemiology of Panton-Valentine Leukocidin-Positive Staphylococcus aureus in Spain: Emergence of the USA300 Clone in an Autochthonous Population. J Clin Microbiol. 2011;49(1):433-436.

Moran GJ, Krishnadasan A, Gorwitz RJ, Gregory E, McDougal LK, Carey RB et al. Methicillin-resistant Staphylococcus aureus infections among patients in the emergency department. N Engl J Med 2006;355:666–674.

Nguyen D, Bancroft E, Mascola L, Guevara R, Yasuda L. Risk factors for neonatal methicillin-resistant Staphylococcus aureus infection in a well-infant nursery. Infect Cont Hosp Epidemiol 2007;28:406–411.

Diep BA, Chambers HF, Graber CJ, Szumowski JD, Miller LG, Han LL, et al. Emergence of multidrug-resistant, community-associated, methicillin-resistant Staphylococcus aureus clone USA 300 in men who have sex with men. Ann Intern Med 2008; 148:249–257.


  • There are currently no refbacks.

Quick links for Authors:Quick links for Peer Reviewers:

Advanced online | Current Issue | Archives | Submit a manuscript | About the Journal

© Association of Basic Medical Sciences of FBIH, 1998-2018. Credits to Public Knowledge Project