Genotyping of Methicillin-resistant Staphylococcus aureus with rep-PCR

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Research Paper 01/06/2017
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Genotyping of Methicillin-resistant Staphylococcus aureus with rep-PCR

Aasma Akhtar, Naima Mahreen, Javaria Altaf, Muhammad Saqib, Mian Abdul Hafeez, Faisal Saeed Awan
Int. J. Biosci.10( 6), 265-275, June 2017.
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Methicillin resistant Staphylococcus aureus (MRSA) is a significant infectious agent which cause infections in human and animals. This study was aimed to genotype human MRSA isolates recovered from wounds. Of the 66 wound cultures, 32 were confirmed as S. aureus and 23 had given positive signal MRSA, respectively. There were consistent banding patterns among all 23 MRSA isolates. The dendrogram attained from the Rep and BOX analysis showed 2 major groups showing a little bit maximum and distant relationship among MRSA strains. Henceforth, this technique could be of valuable use in scheming the modes as well as sources of transmission and mainly inhibiting the hospital related infections those are caused by the MRSA strains.


Aguadero VGonzález Velasco CVindel AGonzalez Velasco MMoreno JJ. 2015. Evaluation of rep-PCR Diversi Lab versus PFGE and spa typing in genotyping methicillin-resistant Staphylococcus aureus (MRSA). Br J Biomed Sci 2015; 72(3), 120-127.

Berger-Bachi B. 2002. Resistance mechanisms of gram-positive bacteria. Int. J. Med. Microbiol 292, 27-35.

Bergey DH, Holt JG, Krieg NR, Sneath PH, Staley JT, Williams ST. 1994. Gram-positive cocci. In, Bergey’s Manual of Determinative Bacteriology, 9th (Ed.). Williams and Wilkins (Eds.). Baltimore, Maryland USA.

Brakstad OG, Assbakk K. Maeland JA. 1992. Detection of Staphylococcus aureus by polymerase chain reaction amplification of the Nuc gene. J. Clin. Microbiol 30, 1600-1654.

Brown DF, Edwards DI, Hawkey PM, Morrison D, Ridgway GL, Towner KJ, Wren MW. 2005. Guidelines for the laboratory diagnosis and susceptibility testing of methicillin-resistant Staphylococcus aureus (MRSA). J. Antimicrob. Chemother 56, 1000-18.

Chambers HF. 2001. The changing epidemiology of Staphylococcus aureus. Emerg. Infect. Dis 7, 178-82.

Del Vecchio VG, Petroziello JM, Gress MJ, McCleskey FK, Melcher GP, Crouch HK, Lupski JR. 1995. Molecular genotyping of methicillin-resistant Staphylococcus aureus via fluorophore-enhanced repetitive-sequence PCR. J. Clin. Microbiol 33, 2141-4.

Enright MC, Robinson DA, Randle G, Feil EJ, Grundmann H. Spratt BG. 2002. The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA). Proc. Natl. Acad. Sci. USA 99, 7687-7692.

Farber JM. 1996. An introduction to the hows and whys of molecular typing. J. Food Prot 59, 1091-101.

Ferry T, Perpoint T, Vandenesch F. Etienne J. 2005. Virulence determinants in Staphylococcus aureus and their involvement in clinical syndromes. Curr. Infect. Dis. Rep 7, 420-8.

Foster TJ. 2009. Colonization and infection of the human host by staphylococci: adhesion, survival and immune evasion. Vet. Dermatol 20, 456-70.

Fournier B. Philpott DJ. 2005. Recognition of Staphylococcus aureus by the innate immune system. Clin. Microbiol. Rev 18, 521-40.

Gordon RJ, Lowy FD. 2008. Pathogenesis of methicillin-resistant Staphylococcus aureus infection. Clin Infect Dis. 2008 Jun 1, 46. Suppl 5: S350-9. DOI: 10.1086/533591.

Grema HA, Geidam YA, Gadzama GB, Ameh JA, Suleiman A. 2015. Methicillin resistant Staphyloccus aureus (MRSA): A review. Adv. Anim. Vet. Sci 3(2), 79-98.

Jeršek B, Tcherneva E, Rijpens N, Herman L. 1996. Repetitive element sequence-based PCR for species and strain discrimination in the genus Listeria. Lett. Appl. Microbiol 23, 55-60.

Jevons MP. 1961. “Celbenin”- Resistant Staphylococci. BMJ 1, 124-5.

Kang HP, Dunne M. 2003. Stability of repetitive sequence PCR patterns with respect to culture age and subculture frequency. J. Clin. Microbiol 41, 2694-2696.

Kwon NH, Park KT, Moon JS, Jung WK, Kim SH, Kim JM, Hong SK, Koo HC, Joo YS, Park YH. 2006. Staphylococcal cassette chromosome mec (SCCmec) characterization and molecular analysis for methicillin-resistant Staphylococcus aureus and novel SCC mec subtype IVg isolated from bovine milk in Korea. J. Antimicrob. Chemother 56, 624-632.

Lee CJ, Sankaran S, Mukherjee DV, Apa ZL, Hafer CA, Wright L, Larson EL, Lowy FD. 2011. Staphylococcus aureus or pharyngeal carriage in a prison population. Clin. Infect. Dis 52, 775-778.

Lowy FD. 1998. Staphylococcus aureus infections. N. Engl. J. Med 339, 520-32.

Mahmood K, Tahir T, Jameel T, Ziauddin A, Aslam HF. 2010. Prevalence of Methicillin-resistant Staphylococcus aureus (MRSA) Causing Nosocomial Infection in a Tertiary Care Hospital. ANNALS 16, 91-96.

Manga I, Vyletělová M. 2012. Rep-PCR-based typing as a tool for tracking of MRSA infection origin. Acta Univ. Agric. Silvic. Mendelianae Brun 60, 251-256.

Marshall JH, Wilmoth GJ. 1981. Pigments of Staphylococcus aureus, a series of triterpenoid carotenoids. J. Bacteriol 147, 900-13.

Maslow JN, Mulligan ME, Arbeit RD. 1993. “Molecular epidemiology: application of contemporary techniques to the typing of microorganisms.” Clin. Infect. Diseases 153-162.

Melzer M, Eykyn SJ, Gransden WR, Chinn S. 2003. Is methicillin resistant Staphylococcus aureus more virulent than methicillin susceptible S. aureus. A comparative cohort study of British patients with nosocomial infection and bacteremia. Clin Infect Dis 37, 1453–60.

Novick RP. 2006. Staphylococcal pathogenesis and pathogenicity factors: Genetics and regulation. In: Fischetti, R.P. J.J. Ferretti, D.A. Portnoy, J.I. Rood. (Eds.). Gram-Positive Pathogens. ASM Press, Washington, D.C 496-516.

Peacock SJ, Paterson GK. 2015. Mechanism of methicillin resistance in Staphylococcus aureus. Annual Rev. Biochem 84, 577-601.

Perwaiz S, Barakzi Q, Farooqi BJ. 2007. Antimicrobial susceptibility pattern of clinical isolates of Methicillin resistant Staphylococcus aureus. J. Pak. Med. Assoc 57, 2-4.

Sabat A, Krzyszton-Russjan J, Strzalka W, Filipek R, Kosowska K, Hryniewicz W, Travis J, Potempa J. 2003. New method for typing Staphylococcus aureus strains: multiple-locus variable-number tandem repeat analysis of polymorphism and genetic relationships of clinical isolates. J. Clin. Microbiol 41, 1801-4.

Saluja SK, Weiser JN. 1995. The genetic basis of colony opacity in Streptococcus pneumoniae: evidence for the effect of box elements on the frequency of phenotypic variation. Mol. Microbiol 16(2), 215-27. 0950-382.

Shu-Hua Wang, Yosef Khan, Lisa Hines, José R, Mediavilla, Liangfen Zhang, Liang Chen, Armando Hoet, Tammy Bannerman, Preeti Pancholi D, Ashley Robinson, Barry N, Kreiswirth, Kurt B, Stevenson. 2012. Methicillin-Resistant Staphylococcus aureus Sequence Type 239-III, Ohio, USA, 2007-2009. Emerg. Infect. Dis 2012 18, L 1557-1565,

Stefani S, Chung DR, Lindsay JA, Friedrich AW, Kearns AM, Westh H, Mackenzie FM. 2012. Meticillin-resistant Staphylococcus aureus (MRSA): global epidemiology and harmonization of typing methods. Int. J. Antimicrob. Agents 39, 273-282.

Stepan J, Pantucek R, Doskar J. 2004. Molecular diagnostics of clinically important staphylococci. Folia Microbiol. (Praha) 49, 353-86.

Swaminathan B, Matar GM. 1993. Molecular typing methods. In: Persing, D.H., T.F. Smith, F.C. Tenover and T.J. White (Eds.). Diagnostic molecular microbiology: principles and applications. Washington 26-50.

Trindade PA, McCulloch JA, Oliveira GA, Mamizuka EM. 2003. Molecular techniques for MRSA typing: current issues and perspectives. Braz. J. Infect. Dis 7, 32-43.

Van Belkum A, Kluitmans J, van leeuwen W, Bax R,  Quint W,  Peters E,  Fluit A,  Vandenbroucke-Grauls C,  van den Brule A,  Koeleman H. 1995. Multicenter evaluation of arbitrarily primed PCR for typing of Staphylococcus aureus strains. J. Clin. Microbiol 33, 1537-47.

Van Belkum A, Sluijuter M, de Groot R, Verbrugh H, Hermans PW. 1996. Novel BOX repeat PCR assay for high-resolution typing of Streptococcus pneumoniae strains. J. Clin. Microbiol 34(5), 1176-9.0095-1137.

Van der Zee A, Verbakel H, van Zon JC, Frenay I,  van Belkum A,  Peeters M,  Buiting A, Bergmans A. 1999. Molecular genotyping of Staphylococcus aureus strains: comparison of repetitive element sequence-based PCR with various typing methods and isolation of a novel epidemicity marker. J. Clin. Microbiol 37, 342-9.

Van Leeuwen W. 2003. Molecular Approaches for Epidemiological Characterization of Staphylococcus aureus. pp 95-136. In. MRSA Current Perspectives. Fluit, A.C. and F.J. Schmitz (Eds.). Caister Academic Press, Norfolk England.

Versalovic J, Kapur V, Koeuth T, Mazurek GH, Whittam TS, Musser JA, Lupski JR. 1994. DNA fingerprinting of pathogenic bacteria by fluorophore-enhanced repetitive sequence-based polymerase chain reaction. Arch. Pathol. Lab. Med 119, 240-246.

Versalovic J, Kapur V, Lupski JR. 1991. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucleic Acids Res 19, 6823-31.

Wieland B, Feil C, Gloria-Maercker E, Thumm G, Lechner M, Bravo JM, Poralla K, Gotz F. 1994. Genetic and biochemical analyses of the biosynthesis of the yellow carotenoid 4, 4’-diaponeurosporene of Staphylococcus aureus. J. Bacteriol 176, 7719-26.

Williams R. 1963. Healthy carriage of Staphylococcus aureus: its prevalence and importance. Bacteriol. Rev 27, 56-71.

Yang A, Yen C. 2012. PCR optimization of BOX-A1R PCR for microbial source tracking of Escherichia coli in waterways. J. Exp. Microbiol. Immunol 16, 85-89.