Hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA) from Pakistan: molecular characterisation by microarray technology

  • Bushra Jamil
  • Darius Gawlik
  • Muhammad Ali Syed
  • Asim Ali Shah
  • Shahid Ahmad Abbasi
  • Elke Müller
  • Annett Reißig
  • Ralf Ehricht
  • Stefan Monecke
Original Article

Abstract

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Pakistan is known to be high, but very few studies have described the molecular epidemiology of the different MRSA clones circulating in the country. Forty-four MRSA isolates were collected from two tertiary care hospitals of the Rawalpindi district of Pakistan. All strains were identified by a conventional phenotypic method and then subjected to genotyping by microarray hybridisation. Six clonal complexes (CCs) and 19 strains were identified. The most commonly identified strains were: (i) Panton–Valentine leucocidin (PVL)-positive CC772-MRSA-V, “Bengal Bay Clone” (ten isolates; 22.3%), (ii) ST239-MRSA [III + ccrC] (five isolates) and (iii) a CC8-MRSA-IV strain, as well as CC6-MRSA-IV (both with four isolates; 9.1% each). Several of the strains detected indicated epidemiological links to the Middle Eastern/Arabian Gulf region. Further studies are needed to type MRSA from countries with less known epidemiology and to monitor the distribution and spread of strains, as well as possible links to global travel, migration and commerce.

Notes

Acknowledgements

Muhammad Ali Syed would like to acknowledge the Vice Chancellor and the Office of Research, Innovation and Commercialization (ORIC) of the University of Haripur for the financial support.

Compliance with ethical standards

Conflict of interest

Darius Gawlik, Elke Müller, Annett Reissig, Ralf Ehricht and Stefan Monecke are employees of Abbott (Alere Technologies GmbH), Jena, the company that manufactures the microarrays used for this study.

Informed consent and ethics approval

Ethic committees consent was not requested since isolates were not purposefully obtained for this study but retrospectively selected after performing routine diagnostics.

Supplementary material

10096_2017_3161_MOESM1_ESM.xlsx (125 kb)
Supplemental file 1 Full hybridisation profiles and additional information on the origin of the isolates. (XLSX 124 kb)

References

  1. 1.
    Ito T, Katayama Y, Asada K, Mori N, Tsutsumimoto K, Tiensasitorn C, Hiramatsu K (2001) Structural comparison of three types of staphylococcal cassette chromosome mec integrated in the chromosome in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 45(5):1323–1336CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Ito T, Ma XX, Takeuchi F, Okuma K, Yuzawa H, Hiramatsu K (2004) Novel type V staphylococcal cassette chromosome mec driven by a novel cassette chromosome recombinase, ccrC. Antimicrob Agents Chemother 48(7):2637–2651CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) (2009) Classification of staphylococcal cassette chromosome mec (SCCmec): guidelines for reporting novel SCCmec elements. Antimicrob Agents Chemother 53(12):4961–4967CrossRefGoogle Scholar
  4. 4.
    Shore AC, Coleman DC (2013) Staphylococcal cassette chromosome mec: recent advances and new insights. Int J Med Microbiol 303(6–7):350-359.  https://doi.org/10.1016/j.ijmm.2013.02.002 CrossRefPubMedGoogle Scholar
  5. 5.
    Deurenberg RH, Vink C, Kalenic S, Friedrich AW, Bruggeman CA, Stobberingh EE (2007) The molecular evolution of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect 13(3):222–235CrossRefPubMedGoogle Scholar
  6. 6.
    Jevons MP (1961) “Celbenin”-resistant staphylococci. Br Med J 1(5219):924–925CrossRefGoogle Scholar
  7. 7.
    Pantosti A, Venditti M (2009) What is MRSA? Eur Respir J 34(5):1190–1196.  https://doi.org/10.1183/09031936.00007709 CrossRefPubMedGoogle Scholar
  8. 8.
    Chambers HF, DeLeo FR (2009) Waves of resistance: Staphylococcus aureus in the antibiotic era. Nat Rev Microbiol 7:629–641CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Coombs GW, Nimmo GR, Bell JM, Huygens F, O’Brien FG, Malkowski MJ, Pearson JC, Stephens AJ, Giffard PM; Australian Group for Antimicrobial Resistance (2004) Genetic diversity among community methicillin-resistant Staphylococcus aureus strains causing outpatient infections in Australia. J Clin Microbiol 42(10):4735–4743CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Enright MC, Day NP, Davies CE, Peacock SJ, Spratt BG (2000) Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. J Clin Microbiol 38(3):1008–1015PubMedPubMedCentralGoogle Scholar
  11. 11.
    Monecke S, Coombs G, Shore AC, Coleman DC, Akpaka P, Borg M, Chow H, Ip M, Jatzwauk L, Jonas D, Kadlec K, Kearns A, Laurent F, O’Brien FG, Pearson J, Ruppelt A, Schwarz S, Scicluna E, Slickers P, Tan H-L, Weber S, Ehricht R (2011) A field guide to pandemic, epidemic and sporadic clones of methicillin-resistant Staphylococcus aureus. PLoS One 6(4):e17936CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Chua K, Laurent F, Coombs G, Grayson ML, Howden BP (2011) Antimicrobial resistance: Not community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA)! A clinician’s guide to community MRSA—its evolving antimicrobial resistance and implications for therapy. Clin Infect Dis 52(1):99–114.  https://doi.org/10.1093/cid/ciq067 CrossRefPubMedGoogle Scholar
  13. 13.
    Otter JA, French GL (2008) The emergence of community-associated methicillin-resistant Staphylococcus aureus at a London teaching hospital, 2000–2006. Clin Microbiol Infect 14(7):670–676CrossRefPubMedGoogle Scholar
  14. 14.
    Miller LG, Perdreau-Remington F, Bayer AS, Diep B, Tan N, Bharadwa K, Tsui J, Perlroth J, Shay A, Tagudar G, Ibebuogu U, Spellberg B (2007) Clinical and epidemiologic characteristics cannot distinguish community-associated methicillin-resistant Staphylococcus aureus infection from methicillin-susceptible S. aureus infection: a prospective investigation. Clin Infect Dis 44(4):471–482CrossRefPubMedGoogle Scholar
  15. 15.
    Monecke S, Jatzwauk L, Müller E, Nitschke H, Pfohl K, Slickers P, Reissig A, Ruppelt-Lorz A, Ehricht R (2016) Diversity of SCCmec elements in Staphylococcus aureus as observed in south-eastern Germany. PLoS One 11(9):e0162654.  https://doi.org/10.1371/journal.pone.0162654 CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Akhter R, Khan K, Hasan F (2009) Isolation and antimicrobial susceptibility pattern of methicillin-resistant and methicillin sensitive Staphylococcus aureus. J Surg Pak 14(4):161–164Google Scholar
  17. 17.
    Ajmal AN, Mir F, Aslam M, Hafeez R (2009) Nosocomial methicillin resistant Staphylococcus aureus frequency in a tertiary care hospital, Lahore, Pakistan. Biomedica 25(2):97–100Google Scholar
  18. 18.
    Butt T, Ahmad RN, Usman M, Mahmood A (2004) Methicillin-resistant Staphylococcus aureus, Pakistan, 1996–2003. Emerg Infect Dis 10(9):1691–1692.  https://doi.org/10.3201/eid1009.030844 CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Hafiz S, Hafiz AN, Ali L, Chughtai AS, Memon B, Ahmed A, Hussain S, Sarwar G, Mughal T, Siddiqui SJ, Awan A, Zaki K, Fareed A (2002) Methicillin resistant Staphylococcus aureus: a multicentre study. J Pak Med Assoc 52(7):312–315PubMedGoogle Scholar
  20. 20.
    Perwaiz S, Barakzi Q, Farooqi BJ, Khursheed N, Sabir N (2007) Antimicrobial susceptibility pattern of clinical isolates of methicillin resistant Staphylococcus aureus. J Pak Med Assoc 57(1):2–4PubMedGoogle Scholar
  21. 21.
    Hussain S, Shams R, Ahmad K, Perveen R, Riaz B (2005) Prevalence of methicillin resistant Staphylococcus aureus (MRSA) in surgical site infections in a tertiary care hospital. J Pathol 3(2):81–85Google Scholar
  22. 22.
    Shabir S, Hardy KJ, Abbasi WS, McMurray CL, Malik SA, Wattal C, Hawkey PM (2010) Epidemiological typing of meticillin-resistant Staphylococcus aureus isolates from Pakistan and India. J Med Microbiol 59(Pt 3):330–337.  https://doi.org/10.1099/jmm.0.014910-0 CrossRefPubMedGoogle Scholar
  23. 23.
    Zafar A, Stone M, Ibrahim S, Parveen Z, Hasan Z, Khan E, Hasan R, Wain J, Bamford K (2011) Prevalent genotypes of meticillin-resistant Staphylococcus aureus: report from Pakistan. J Med Microbiol 60(Pt 1):56–62.  https://doi.org/10.1099/jmm.0.022707-0 CrossRefPubMedGoogle Scholar
  24. 24.
    Madzgalla S, Syed MA, Khan MA, Rehman SS, Müller E, Reissig A, Ehricht R, Monecke S (2016) Molecular characterization of Staphylococcus aureus isolates causing skin and soft tissue infections in patients from Malakand, Pakistan. Eur J Clin Microbiol Infect Dis 35(9):1541–1547.  https://doi.org/10.1007/s10096-016-2695-8 CrossRefPubMedGoogle Scholar
  25. 25.
    Afroz S, Kobayashi N, Nagashima S, Alam MM, Hossain AB, Rahman MA, Islam MR, Lutfor AB, Muazzam N, Khan MA, Paul SK, Shamsuzzaman AK, Mahmud MC, Musa AK, Hossain MA (2008) Genetic characterization of Staphylococcus aureus isolates carrying Panton–Valentine leukocidin genes in Bangladesh. Jpn J Infect Dis 61(5):393–396PubMedGoogle Scholar
  26. 26.
    Blomfeldt A, Larssen KW, Moghen A, Gabrielsen C, Elstrøm P, Aamot HV, Jørgensen SB (2017) Emerging multidrug-resistant Bengal Bay clone ST772-MRSA-V in Norway: molecular epidemiology 2004–2014. Eur J Clin Microbiol Infect Dis 36(10):1911–1921.  https://doi.org/10.1007/s10096-017-3014-8 CrossRefPubMedGoogle Scholar
  27. 27.
    Brennan GI, Shore AC, Corcoran S, Tecklenborg S, Coleman DC, O’Connell B (2012) Emergence of hospital- and community-associated Panton–Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus genotype ST772-MRSA-V in Ireland and detailed investigation of an ST772-MRSA-V cluster in a neonatal intensive care unit. J Clin Microbiol 50(3):841–847.  https://doi.org/10.1128/jcm.06354-11 CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Ellington MJ, Ganner M, Warner M, Cookson BD, Kearns AM (2010) Polyclonal multiply antibiotic-resistant methicillin-resistant Staphylococcus aureus with Panton–Valentine leucocidin in England. J Antimicrob Chemother 65(1):46–50CrossRefPubMedGoogle Scholar
  29. 29.
    Mediavilla JR, Chen L, Mathema B, Kreiswirth BN (2012) Global epidemiology of community-associated methicillin resistant Staphylococcus aureus (CA-MRSA). Curr Opin Microbiol 15(5):588–595.  https://doi.org/10.1016/j.mib.2012.08.003 CrossRefPubMedGoogle Scholar
  30. 30.
    Monecke S, Baier V, Coombs GW, Slickers P, Ziegler A, Ehricht R (2013) Genome sequencing and molecular characterisation of Staphylococcus aureus ST772-MRSA-V, “Bengal Bay Clone”. BMC Res Notes 6(1):548.  https://doi.org/10.1186/1756-0500-6-548 CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Prabhakara S, Khedkar S, Shambat SM, Srinivasan R, Basu A, Norrby-Teglund A, Seshasayee ASN, Arakere G (2013) Genome sequencing unveils a novel sea enterotoxin-carrying PVL phage in Staphylococcus aureus ST772 from India. PLoS One 8(3):e60013.  https://doi.org/10.1371/journal.pone.0060013 CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Sanchini A, Campanile F, Monaco M, Cafiso V, Rasigade JP, Laurent F, Etienne J, Stefani S, Pantosti A (2011) DNA microarray-based characterisation of Panton–Valentine leukocidin-positive community-acquired methicillin-resistant Staphylococcus aureus from Italy. Eur J Clin Microbiol Infect Dis 30(11):1399–1408.  https://doi.org/10.1007/s10096-011-1234-x CrossRefPubMedGoogle Scholar
  33. 33.
    Shore AC, Tecklenborg SC, Brennan GI, Ehricht R, Monecke S, Coleman DC (2014) Panton–Valentine leukocidin-positive Staphylococcus aureus in Ireland from 2002 to 2011: 21 clones, frequent importation of clones, temporal shifts of predominant methicillin-resistant S. aureus clones, and increasing multiresistance. J Clin Microbiol 52(3):859–870.  https://doi.org/10.1128/JCM.02799-13 CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Udo EE, Al-Lawati BAH, Al-Muharmi Z, Thukral SS (2014) Genotyping of methicillin-resistant Staphylococcus aureus in the Sultan Qaboos University Hospital, Oman reveals the dominance of Panton–Valentine leucocidin-negative ST6-IV/t304 clone. New Microbes New Infect 2(4):100–105.  https://doi.org/10.1002/nmi2.47 CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Williamson DA, Roberts SA, Ritchie SR, Coombs GW, Fraser JD, Heffernan H (2013) Clinical and molecular epidemiology of methicillin-resistant Staphylococcus aureus in New Zealand: rapid emergence of sequence type 5 (ST5)-SCCmec-IV as the dominant community-associated MRSA clone. PLoS One 8(4):e62020.  https://doi.org/10.1371/journal.pone.0062020 CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Monecke S, Slickers P, Ehricht R (2008) Assignment of Staphylococcus aureus isolates to clonal complexes based on microarray analysis and pattern recognition. FEMS Immunol Med Microbiol 53:237–251CrossRefPubMedGoogle Scholar
  37. 37.
    Scicluna EA, Shore AC, Thürmer A, Ehricht R, Slickers P, Borg MA, Coleman DC, Monecke S (2010) Characterisation of MRSA from Malta and the description of a Maltese epidemic MRSA strain. Eur J Clin Microbiol Infect Dis 29(2):163–170CrossRefPubMedGoogle Scholar
  38. 38.
    Monecke S, Skakni L, Hasan R, Ruppelt A, Ghazal SS, Hakawi A, Slickers P, Ehricht R (2012) Characterisation of MRSA strains isolated from patients in a hospital in Riyadh, Kingdom of Saudi Arabia. BMC Microbiol 12(1):146CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Coombs G, Pearson J, Daley D, Robinson O, Nimmo G, Turnidge JD (2013) Staphylococcus aureus Programme 2012 (SAP 2012) community survey. MRSA epidemiology and typing report. Australian Group on Antimicrobial Resistance (AGAR), AustraliaGoogle Scholar
  40. 40.
    Holden MTG, Hsu L-Y, Kurt K, Weinert LA, Mather AE, Harris SR, Strommenger B, Layer F, Witte W, de Lencastre H, Skov R, Westh H, Žemličková H, Coombs G, Kearns AM, Hill RLR, Edgeworth J, Gould I, Gant V, Cooke J, Edwards GF, McAdam PR, Templeton KE, McCann A, Zhou Z, Castillo-Ramírez S, Feil EJ, Hudson LO, Enright MC, Balloux F, Aanensen DM, Spratt BG, Fitzgerald JR, Parkhill J, Achtman M, Bentley SD, Nübel U (2013) A genomic portrait of the emergence, evolution, and global spread of a methicillin-resistant Staphylococcus aureus pandemic. Genome Res 23(4):653–664.  https://doi.org/10.1101/gr.147710.112 CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Robinson DA, Enright MC (2004) Evolution of Staphylococcus aureus by large chromosomal replacements. J Bacteriol 186(4):1060–1064CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Biber A, Abuelaish I, Rahav G, Raz M, Cohen L, Valinsky L, Taran D, Goral A, Elhamdany A, Regev-Yochay G; PICR Study Group (2012) A typical hospital-acquired methicillin-resistant Staphylococcus aureus clone is widespread in the community in the Gaza strip. PLoS One 7(8):e42864.  https://doi.org/10.1371/journal.pone.0042864 CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Al Laham N, Mediavilla JR, Chen L, Abdelateef N, Elamreen FA, Ginocchio CC, Pierard D, Becker K, Kreiswirth BN (2015) MRSA clonal complex 22 strains harboring toxic shock syndrome toxin (TSST-1) are endemic in the primary Hospital in Gaza, Palestine. PLoS One 10(3):e0120008.  https://doi.org/10.1371/journal.pone.0120008 CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Udo EE, Boswihi SS, Al-Sweih N (2016) High prevalence of toxic shock syndrome toxin-producing epidemic methicillin-resistant Staphylococcus aureus 15 (EMRSA-15) strains in Kuwait hospitals. New Microbes New Infect 12:24–30.  https://doi.org/10.1016/j.nmni.2016.03.008 CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Senok A, Somily A, Raji A, Gawlik D, Al-Shahrani F, Baqi S, Boswihi S, Skakni L, Udo EE, Weber S, Ehricht R, Monecke S (2016) Diversity of methicillin-resistant Staphylococcus aureus CC22-MRSA-IV from Saudi Arabia and the Gulf region. Int J Infect Dis 51:31–35CrossRefPubMedGoogle Scholar
  46. 46.
    Monecke S, Aamot HV, Stieber B, Ruppelt A, Ehricht R (2014) Characterization of PVL-positive MRSA from Norway. APMIS 122(7):580–584.  https://doi.org/10.1111/apm.12181 CrossRefPubMedGoogle Scholar
  47. 47.
    Harris SR, Feil EJ, Holden MT, Quail MA, Nickerson EK, Chantratita N, Gardete S, Tavares A, Day N, Lindsay JA, Edgeworth JD, de Lencastre H, Parkhill J, Peacock SJ, Bentley SD (2010) Evolution of MRSA during hospital transmission and intercontinental spread. Science 327(5964):469–474CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Chen H, Yang X, Wang Q, Zhao C, Li H, He W, Wang X, Zhang F, Wang Z, Chen M, Zhu B, Wang H (2014) Insights on evolution of virulence and resistance from the whole-genome analysis of a predominant methicillin-resistant Staphylococcus aureus clone sequence type 239 in China. Chin Sci Bull 59(11):1104–1112.  https://doi.org/10.1007/s11434-014-0149-1 CrossRefGoogle Scholar
  49. 49.
    Yamamoto T, Takano T, Higuchi W, Iwao Y, Singur O, Reva I, Otsuka Y, Nakayashiki T, Mori H, Reva G, Kuznetsov V, Potapov V (2012) Comparative genomics and drug resistance of a geographic variant of ST239 methicillin-resistant Staphylococcus aureus emerged in Russia. PLoS One 7(1):e29187.  https://doi.org/10.1371/journal.pone.0029187 CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    Weber S, Ehricht R, Slickers P, Abdel-Wareth L, Donnelly G, Pitout M, Monecke S (2010) Genetic fingerprinting of MRSA from Abu Dhabi. In: Proceedings of the 20th European Congress of Clinical Microbiology and Infectious Diseases (ECCMID), Vienna, Austria, April 2010Google Scholar
  51. 51.
    Witte W, Enright M, Schmitz FJ, Cuny C, Braulke C, Heuck D (2001) Characteristics of a new epidemic MRSA in Germany ancestral to United Kingdom EMRSA 15. Int J Med Microbiol 290(8):677–682CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  • Bushra Jamil
    • 1
    • 2
  • Darius Gawlik
    • 3
    • 4
  • Muhammad Ali Syed
    • 5
  • Asim Ali Shah
    • 6
  • Shahid Ahmad Abbasi
    • 7
  • Elke Müller
    • 3
    • 4
  • Annett Reißig
    • 3
    • 4
  • Ralf Ehricht
    • 3
    • 4
  • Stefan Monecke
    • 3
    • 4
    • 8
  1. 1.Department of BiosciencesCOMSATS Institute of Information TechnologyIslamabadPakistan
  2. 2.National University of Medical Sciences (NUMS)RawalpindiPakistan
  3. 3.Abbott (Alere Technologies GmbH)JenaGermany
  4. 4.InfectoGnostics Research CampusJenaGermany
  5. 5.Department of MicrobiologyUniversity of HaripurHaripurPakistan
  6. 6.Department of PathologyFauji Foundation HospitalRawalpindiPakistan
  7. 7.Al-Sayed Hospital (Pvt) Ltd.RawalpindiPakistan
  8. 8.Institute for Medical Microbiology and HygieneTechnische Universität DresdenDresdenGermany

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