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:: Volume 11, Issue 3 (6-2024) ::
2024, 11(3): 35-45 Back to browse issues page
Spa typing of Methicillin-Resistant Staphylococcus aureus isolated from clinical samples of hospitalized patients, a study in the Wasit province of Iraq
Karar Alhakeem , Mostafa Nemati , Fazel Pourahmad , Hussam Sami Alshimry
Associate Prof., Department of Microbiology, Faculty of Veterinary Sciences, Ilam University, Ilam-Iran , m.nemati@ilam.ac.ir
Abstract:   (98 Views)
Introduction: Since its discovery in 1961, methicillin-resistant Staphylococcus aureus (MRSA) has been recognized as a significant healthcare-associated pathogen (HA-MRSA) and a notorious 'superbug'. Typing is crucial for surveillance, epidemiology analysis, infection control of MRSA and sequencing of the spa gene is one of the most common methods used for determining the origin of this bacterium in humans and animals. This research aimed to determine the antibiotic resistance and spa type of S. aureus strains collected from outpatients in two hospitals in the Wasit province of Iraq.
Material & Methods: The study analyzed 200 outpatient MRSA isolates by collecting nasal and sputum samples from patients. Standard biochemical and molecular methods based on the nuc gene were used to identify S. aureus bacteria and amplify the mecA and spa genes. The Kirby-Bauer disc diffusion method was employed to determine the antibiotic sensitivity of the isolates using penicillin, cefoxitin, vancomycin, gentamicin, erythromycin, tetracycline, imipenem, clindamycin, chloramphenicol and rifampicin.
Results: Thirty-five (17.5%) out of 200 isolates were identified as S. aureus by biochemical and molecular methods. The prevalence of MRSA was more common in women than in men. Antibiogram results showed that most of the isolates were resistant to penicillin (94.2%) and sensitive to imipenem (100%), clindamycin (100%), and chloramphenicol (100%). Of these 35 isolates, 30 (87.5%) and 26 strains (74.3%) were positive for the mecA and spa genes. Typing based on spa gene sequencing revealed four different patterns: t386, t3579, t10002 and t10234.
Conclusion:  Variations in the spa gene among different S. aureus isolates may be of clinical importance when treating staphylococcal infections. In this study, spa typing revealed four different patterns in Iraq, representing diagnostic and therapeutic implications
Keywords: : Staphylococcus aureus, MRSA, PCR, mecA, spa typing
Full-Text [PDF 1831 kb]   (31 Downloads)    
Type of Study: Research | Subject: Clinical microbiology
Received: 2024/02/13 | Accepted: 2024/05/21 | Published: 2024/06/21
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47. Robinson DA, Enright MC. Multilocus sequence typing and the evolution of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect. 2004;10(2):92-7. [DOI:10.1111/j.1469-0691.2004.00768.]
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51. Harmsen D, Claus H, Witte W, Rothganger J, Claus H, Turnwald D, Vogel U. 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(12):5442-8. [DOI:10.1128/JCM.41.12.5442–5448.2003.]
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53. Thaker HC, Brahmbhatt MN, Nayak JB, Thaker HC. Isolation and identification of Staphylococcus aureus from milk and milk products and their drug resistance patterns in Anand, Gujarat. Vet World. 2013;6(1):10-3. https://doi:10.5455/vetworld.2013.10-13.
54. Flayhart D, Lema C, Borek A, Carroll KC. Comparison of the BBL CHROMagar Staph aureus agar medium to conventional media for detection of Staphylococcus aureus in respiratory samples. J Clin Microbiol. 2004;42(8):3566-9. [DOI:10.1128/jcm.42.8.3566-3569.2004.]
55. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; 28th informational supplement, M100-S8. Clinical and Laboratory Standards Institute, Wayne, PA. 2018.
56. Kateete DP, Kimani CN, Katabazi FA, Okeng A, Okee MS, Nanteza A, Joloba ML, Najjuka FC. Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test. Ann. Clin Microbiol Antimicrob. 2010;9:1-7. [DOI:10.1186/1476-0711-9-23.]
57. Al-Talib H, Yean CY, Al-Khateeb A, Hassan H, Singh KK, Al-Jashamy K, Ravichandran M. A pentaplex PCR assay for the rapid detection of methicillin-resistant Staphylococcus aureus and Panton-Valentine Leucocidin. BMC Microbiology. 2009;9:1-8. doi.org/10.1186/1471-2180-9-113
58. Wichelhaus TA, Hunfeld KP, Böddinghaus B, Kraiczy P, Schafer V, Brade V. Rapid molecular typing of methicillin-resistant Staphylococcus aureus by PCR-RFLP. Infection Control & Hospital Epidemiology. 2001;22(5):294-8. [DOI:10.1086/501903.]
59. Al-Mayahi A, Srhan F. A preliminary study of Aminoglycoside Modifying Enzymes (AMEs) of Multiple Antibiotic Resistance of Methicillin-resistant Staphylococcus aureus (MRSA) isolated from clinical specimens in Al-Diwaniya/Iraq. Jordan J Biol Sci. 2021;14(4).733-41.
60. Saleh NT, Alsammak eg. spa typing and virulence genes variation in Methicillin -resistant Staphylococcus aureus isolated from Mosul, Iraq. Biochem Cell Arch. 2022;22(1). connectjournals.com/03896.2022.22.2163.
61. Al-Charrakh AH, Al-Hassnawi HH, Al-Khafaji JK. Molecular characteristics of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) isolates from clinical specimens in Iraq. Br Microbiol Res J. 2015;5(3):227. [DOI:10.9734/BMRJ/2015/9607.]
62. Hallabjaiy RM, Darogha SN, Hamad PA. Vancomycin resistance among methicillin-resistant Staphylococcus aureus isolated from clinical samples in Erbil City-Iraq. Medical Journal of Islamic World Acad Sci. 2014;22(4):168-74.
63. János D, Viorel H, Ionica I, Corina P, Tiana F, Roxana D. Carriage of multidrug resistance staphylococci in shelter dogs in Timisoara, Romania. Antibiotics. 2021;10(7):801. [DOI:10.3390/antibiotics10070801.]
64. El-Adawy H, Ahmed M, Hotzel H, Monecke S, Schulz J, Hartung J, Ehricht R, Neubauer H, Hafez HM. Characterization of methicillin-resistant Staphylococcus aureus isolated from healthy turkeys and broilers using DNA microarrays. Front Microbiol. 2016;19;7:2019. [DOI:10.3389/fmicb.2016.02019.]
65. Hiramatsu K, Hanaki H, Ino T, Yabuta K, Oguri T, Tenover FC. Methicillin-resistant Staphylococcus aureus clinical strain with reduced vancomycin susceptibility. J Antimicrob Chemother. 1997;40(1):135-6.
66. Liu J, Gao Y, Wang X, Qian Z, Chen J, Huang Y, Meng Z, Lu X, Deng G, Liu F, Zhang Z. Culture-positive spontaneous ascitic infection in patients with acute decompensated cirrhosis: multidrug-resistant pathogens and antibiotic strategies. Yonsei Med J. 2020;61(2):145-53. [DOI:10.3349/ymj.2020.61.2.14.]
67. Mwansa TN, Kamvuma K, Mulemena JA, Phiri CN, Chanda W. Antibiotic susceptibility patterns of pathogens isolated from laboratory specimens at Livingstone Central Hospital in Zambia. PLoS One. 2022;2(9):e0000623. [DOI:10.1371/journal.pgph.0000623.]
68. Yehia HM, Al-Masoud AH, Alarjani KM, Alamri MS. Prevalence of methicillin-resistant (mecA gene) and heat-resistant Staphylococcus aureus strains in pasteurized camel milk. J Dairy Sci. 2020;103(7):5947-63. [DOI:10.3168/jds.2019-17631.]
69. Goering RV, Swartzendruber EA, Obradovich AE, Tickler IA, Tenover FC. Emergence of oxacillin resistance in stealth methicillin-resistant Staphylococcus aureus due to mecA sequence instability. Antimicrob Agents Chemother. 2019;63(8):10-128. [DOI:10.1128/aac.00558-19.]
70. Jannati E, Arzanlou M, Habibzadeh S, Mohammadi S, Ahadi P, Mohammadi-Ghalehbin B, Dogaheh HP, Dibah S, Kazemi E. Nasal colonization of mecA-positive, oxacillin-susceptible, methicillin-resistant Staphylococcus aureus isolates among nursing staff in an Iranian teaching hospital. Am J Infect Control. 2013;41(11):1122-4. [DOI:10.1016/j.ajic.2013.02.012.]
71. Chung M, Kim CK, Conceiçao T, Aires-De-Sousa M, De Lencastre H, Tomasz A. Heterogeneous oxacillin-resistant phenotypes and production of PBP2A by oxacillin-susceptible/mecA-positive MRSA strains from Africa. J Antimicrob Chemother. 2016;71(10):2804-9. [DOI:10.1093/jac/dkw209.]
72. Duarte FC, Danelli T, Tavares ER, Morguette AE, Kerbauy G, Grion CM, Yamauchi LM, Perugini MR, Yamada-Ogatta SF. Fatal sepsis caused by mecA-positive oxacillin-susceptible Staphylococcus aureus: first report in a tertiary hospital of southern Brazil. J Infect Chemother. 2019;25(4):293-7. [DOI:10.1016/j.jiac.2018.09.010.]
73. Gargis AS, Yoo BB, Lonsway DR, Anderson K, Campbell D, Ewing TO, Lawsin A, Machado MJ, Yamamoto N, Halpin AL, Lutgring JD. Difficult-to-detect Staphylococcus aureus: mecA-positive isolates associated with oxacillin and cefoxitin false-susceptible results. J. Clin Microbiol. 2020;58(4):10-128. https://journals.asm.org/doi/pdf/10.1128/jcm.02038-19.
74. Baddour MM, AbuElKheir MM, Fatani AJ. Comparison of mecA polymerase chain reaction with phenotypic methods for the detection of methicillin-resistant Staphylococcus aureus. Curr Microbiol. 2007;55: 473-9. [DOI:10.1007/s00284-007-9015-6.]
75. H Al-Hassnawi H, H Al-Charrakh A, Al-Khafaj J. Antibiotic resistance patterns of community acquired methicillin resistance Staphylococcus aureus (CA-MRSA) in Al-Hilla/Iraq. Karbala J Pharmaceut Sci. 2013;4(4):91-102.
76. Ba X, Harrison EM, Edwards GF, Holden MT, Larsen AR, Petersen A, Skov RL, Peacock SJ, Parkhill J, Paterson GK, Holmes MA. Novel mutations in penicillin-binding protein genes in clinical Staphylococcus aureus isolates that are methicillin resistant on susceptibility testing, but lack the mec gene. J Antimicrob Chemother. 2014;69(3):594-7. [DOI:10.1093/jac/dkt418.]
77. Keener AB, Thurlow LT, Kang S, Spidale NA, Clarke SH, Cunnion KM, Tisch R, Richardson AR, Vilen BJ. Staphylococcus aureus protein A disrupts immunity mediated by long-lived plasma cells. J Immunol. 2017 1;198(3):1263-73. [DOI:10.4049/jimmunol.1600093.]
78. Tahoun A, Elnafarawy HK, El-Sharkawy H, Rizk AM, Alorabi M, El-Shehawi AM, Youssef MA, Ibrahim HM, El-Khodery S. The prevalence and molecular biology of Staphylococcus aureus isolated from healthy and diseased equine eyes in Egypt. Antibiotics. 2022;10;11(2):221. [DOI:10.3390/antibiotics11020221.]
79. Shakeri F, Shojai A, Golalipour M, Rahimi Alang S, Vaez H, Ghaemi EA. Spa Diversity among MRSA and MSSA Strains of Staphylococcus aureus in North of Iran. Int J Microbiol. 2010; 2010:1-5. [DOI:10.1155/2010/351397.]
80. Al-Kadmy IM. A genetic study to differential HC/AC MRSA isolated from clinical cases in Iraq hospitals. Mintage J Pharm Med Sci. 2013; 2:57-62.
81. Craft KM, Nguyen JM, Berg LJ, Townsend SD. Methicillin-resistant Staphylococcus aureus (MRSA): antibiotic-resistance and the biofilm phenotype. Med Chem Comm. 2019;10(8):1231-41. [DOI:10.1039/C9MD00044E.]
82. Kareem SM, Aljubori SS, Ali MR. Novel determination of spa gene diversity and its molecular typing among Staphylococcus aureus Iraqi isolates obtained from different clinical samples. New Microbes New Infect. 2020; 34:100653. [DOI:10.1016/j.nmni.2020.100653.]
83. Adwan G, Shaheen H, Adwan K, Barakat A. Molecular characterization of methicillin resistant Staphylococcus aureus isolated from hospitals environments and patients in Northern Palestine. Epidemiol Biostat Public Health. 2015;12(3). [DOI:10.2427/11183.]
84. Aqel AA, Alzoubi HM, Vickers A, Pichon B, Kearns AM. Molecular epidemiology of nasal isolates of methicillin-resistant Staphylococcus aureus from Jordan. J Infect Public Health. 2015;8(1):90-7. [DOI:10.1016/j.jiph.2014.05.007.]
85. Harastani HH, Araj GF, Tokajian ST. Molecular characteristics of Staphylococcus aureus isolated from a major hospital in Lebanon. Int J Infect Dis. 2014; 19:33-8. [DOI:10.1016/j.ijid.2013.10.007.]
86. Goudarzi M, Fazeli M, Goudarzi H, Azad M, Seyedjavadi SS. spa typing of Staphylococcus aureus strains isolated from clinical specimens of patients with nosocomial infections in Tehran, Iran. Jundishapur J Microbiol. 2016; 9 (7): 1-9. [DOI:10.5812%2Fjjm.35685.]
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Alhakeem K, Nemati M, Pourahmad F, Sami Alshimry H. Spa typing of Methicillin-Resistant Staphylococcus aureus isolated from clinical samples of hospitalized patients, a study in the Wasit province of Iraq. Journal of Basic Research in Medical Sciences 2024; 11 (3) :35-45
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Volume 11, Issue 3 (6-2024) Back to browse issues page
مجله ی تحقیقات پایه در علوم پزشکی Journal of Basic Research in Medical Sciences
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