Molecular typing of Methicillin Resistance Staphylococcus aureus(MRSA) isolated from device related infections by SCCmec and PCR-RFLP of coagulase gene

Muhamad Sohail, Zakia Latif


Background: SCCmec and PCR-RFLP are productive and cost-effective methods for epidemiological investigations and source tracking of MRSA. Aim of this study was to investigate the epidemiology and molecular characterization of MRSA recovered from device related infection. Methods: A total of 626 MRSA were collected from prosthetic device related infections and subjected for presence of mec gene and SCCmec typing. SCCmec characterized MRSA were subjected for agr typing followed by RFLP genotyping and amplification of PVL gene. Following the Clinical & Laboratory Standards Institute (CLSI), antibiotic resistance pattern was determined. Results: Of 626 MRSA isolates, 488(78%) were characterized by SCCmec typing. Most common type was SCCmec IV (43%), followed by SCCmec II (25%) SCCmec III (22%) and only 10% was SCCmec V.  SCCmec characterized MRSA strains were 100% resistance to tobramycin and chloramphenicol, 96% resistance to ciprofloxacin and 93% resistance to azithromycin. The genePVL was only present in SCCmec IV. All agr typed MRSA strains were resistance to gentamicin, tobramycin and chloramphenicol. PVL was present in all strains of agr III, 80% of agrIV, 50% of agrII and absent in agrI. RFLP analysis resulted in 16 types of non-duplicate unique bands pattern which were equally distributed among prosthetic device related infection. Conclusion: PVL harboring SCCmec or agr typed MRSA strains are less resistance to antibiotics. RFLP is simple, productive and cost-effective method for molecular typing of MRSA. Correct use of antibiotics and molecular surveillance is indispensable to detect the change in epidemiological and antibiotic resistance trends of MRSA.

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Zimmerli W, Trampuz A, Ochsner PE. Prosthetic-joint infections. New England Journal of Medicine, (2004); 351(16): 1645-1654.

CLSI (2015) M100-S26. Performance Standards for Antimicrobial Susceptibility Testing: 26th Informational supplement. Clinical and Laboratory Standards Institute Wayne, PA.

Peerayeh SN, Azimian A, Nejad QB, Kashi M. Prevalence of agr specificity groups amongStaphylococcus aureus isolates from university hospitals in Tehran. Laboratory Medicine, (2015); 40(1): 27-29.

Wright JS, Jin R, Novick RP. Transient interference with staphylococcal quorum sensing blocks abscess formation. Proceedings of the National Academy of Sciences of the United States of America, (2005); 102(5): 1691-1696.

Wong H, Louie L, Lo RY, Simor AE. Characterization of Staphylococcus aureus isolates with a partial or complete absence of staphylococcal cassette chromosome elements. Journal of clinical microbiology, (2010); 48(10): 3525-3531.

Arakere G, Nadig S, Swedberg G, Macaden R, Amarnath SK, et al. Genotyping of methicillin-resistant Staphylococcus aureus strains from two hospitals in Bangalore, South India. Journal of clinical microbiology, (2005); 43(7): 3198-3202.

DeLeo FR, Diep BA, Otto M. Host defense and pathogenesis in Staphylococcus aureusinfections. Infectious Disease Clinics, (2009); 23(1): 17-34.

Sohail M, Latif Z. Prevalence and antibiogram of methicillin resistant Staphylococcus aureusisolated from medical device-related infections; a retrospective study in Lahore, Pakistan. Revista da Sociedade Brasileira de Medicina Tropical, (2017); 50(5): 680-684.

Wilson K. Preparation of genomic DNA from bacteria. Curr Protoc Mol Biol, (2001); Chapter 2Unit 2.4.

Makgotlho PE, Kock MM, Hoosen A, Lekalakala R, Omar S, et al. Molecular identification and genotyping of MRSA isolates. FEMS Immunology & Medical Microbiology, (2009); 57(2): 104-115.

McClure J-A, Conly JM, Lau V, Elsayed S, Louie T, et al. Novel multiplex PCR assay for detection of the staphylococcal virulence marker Panton-Valentine leukocidin genes and simultaneous discrimination of methicillin-susceptible from-resistant staphylococci. Journal of clinical microbiology, (2006); 44(3): 1141-1144.

Nelson MU, Bizzarro MJ, Baltimore RS, Dembry LM, Gallagher PG. Clinical and molecular epidemiology of methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit in the decade following implementation of an active detection and isolation program. Journal of clinical microbiology, (2015); 53(8): 2492-2501.

Himabindu M, Muthamilselvan DS, Bishi DK, Verma RS. Molecular analysis of coagulase gene polymorphism in clinical isolates of methicillin resistant Staphylococcus aureus by restriction fragment length polymorphism based genotyping. American Journal of Infectious Diseases, (2009); 5170-176.

Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution, (1985); 39(4): 783-791.

Sohail M, Latif Z. Prevalence and antibiogram of methicillin resistant Staphylococcus aureusisolated from medical device-related infections; a retrospective study in Lahore, Pakistan. Revista da Sociedade Brasileira de Medicina Tropical, (2017); 50: 680-684.

Havaei SA, Ghanbari F, Rastegari AA, Azimian A, Khademi F, et al. Molecular typing of hospital-acquired Staphylococcus aureus isolated from Isfahan, Iran. International scholarly research notices, (2014); 185272.

Weller T. Methicillin-resistant Staphylococcus aureus typing methods: which should be the international standard. Journal of Hospital Infection, (2000); 44(3): 160-172.

Nasirian S, Saadatmand S, Goudarzi M. Molecular typing of methicillin-resistantStaphylococcus aureus strains isolated from patients admitted to intensive care units of hospitals in Tehran city, Iran, based on agr pattern. Journal of Isfahan Medical School, (2017); 35(451): 1424-1431.

Bayat B, Zade MH, Mansouri S, Kalantar E, Kabir K, et al. High frequency of methicillin-resistant Staphylococcus aureus (MRSA) with SCCmec type III and spa type t030 in Karaj's teaching hospitals, Iran. Acta Microbiologica et Immunologica Hungarica, (2017); 64(3): 331-341.

Perovic O, Singh-Moodley A, Govender NP, Kularatne R, Whitelaw A, et al. A small proportion of community-associated methicillin-resistant Staphylococcus aureus bacteraemia, compared to healthcare-associated cases, in two South African provinces. European Journal of Clinical Microbiology and Infectious Diseases, (2017); 36(12): 2519-2532.

Udo EE, Boswihi SS. Antibiotic Resistance Trends in Methicillin-Resistant Staphylococcus aureus Isolated in Kuwait Hospitals: 2011-2015. Medical Principles and Practice, (2017); 26(5): 485-490.

Vali L, Dashti AA, Mathew F, Udo EE. Characterization of heterogeneous MRSA and MSSA with Reduced Susceptibility to chlorhexidine in Kuwaiti hospitals. Frontiers in Microbiology, (2017); 8(JUL).

Eed EM, Ghonaim MM, Hussein YM, Saber TM, Khalifa AS. Phenotypic and molecular characterization of HA-MRSA in Taif hospitals, Saudi Arabia. The Journal of Infection in Developing Countries, (2015); 9(03): 298-303.

Nada T, Ichiyama S, Osada Y, Ohta M, Shimokata K, et al. Comparison of DNA fingerprinting by PFGE and PCR-RFLP of the coagulase gene to distinguish MRSA isolates. Journal of Hospital Infection, (1996); 32(4): 305-317.

Goudarzi H, Seyedjavadi SS, Udo EE, Beiranvand E, Fazeli M, et al. Molecular Characterization and Distribution of Class 1 Integron-Bearing Methicillin ResistantStaphylococcus aureus Strains in Burn Patients, Tehran, Iran. Jundishapur Journal of Microbiology, (2017); 10(2): e40592.

Lu Z, Chen Y, Chen W, Liu H, Song Q, et al. Characteristics of qacA/B-positive Staphylococcus aureus isolated from patients and a hospital environment in China. Journal of Antimicrobial Chemotherapy, (2014); 70(3): 653-657.

Khemiri M, Alhusain AA, Abbassi MS, El Ghaieb H, Costa SS, et al. Clonal spread of methicillin-resistant Staphylococcus aureus-t6065-CC5-SCCmecV-agrII in a Libyan hospital. Journal of global antimicrobial resistance, (2017); 10: 101-105.

Mohammadi S, Sekawi Z, Monjezi A, Maleki M-H, Soroush S, et al. Emergence of SCCmec type III with variable antimicrobial resistance profiles and spa types among methicillin-resistantStaphylococcus aureus isolated from healthcare-and community-acquired infections in the west of Iran. International Journal of Infectious Diseases, (2014); 25: 152-158.

Bhatta DR, Cavaco LM, Nath G, Kumar K, Gaur A, et al. Association of Panton Valentine Leukocidin (PVL) genes with methicillin resistant Staphylococcus aureus (MRSA) in Western Nepal: a matter of concern for community infections (a hospital based prospective study). BMC infectious diseases, (2016); 16(1): 199.

Qin Y, Wen F, Zheng Y, Zhao R, Hu Q, et al. Antimicrobial resistance and molecular characteristics of methicillin-resistant Staphylococcus aureus isolates from child patients of high-risk wards in Shenzhen, China. Japanese journal of infectious diseases, (2017); 70(5): 479-484.

Monecke S, Skakni L, Hasan R, Ruppelt A, Ghazal SS, et al. Characterisation of MRSA strains isolated from patients in a hospital in Riyadh, Kingdom of Saudi Arabia. BMC microbiology, (2012); 12(1): 146.

Udo EE, Al-Lawati B-H, Al-Muharmi Z, Thukral S. Genotyping of methicillin-resistantStaphylococcus aureus in the Sultan Qaboos University Hospital, Oman reveals the dominance of Panton–Valentine leucocidin-negative ST6-IV/t304 clone. New microbes and new infections, (2014); 2(4): 100-105.


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