Molecular characterization of typing and subtyping of Staphylococcal cassette chromosome SCCmec types I to V in methicillin-resistant Staphylococcus aureus from clinical isolates from COVID-19 patients
Abstract
Background and Objectives: Methicillin resistance is acquired by the bacterium due to mecA gene which codes for penicillin-binding protein (PBP2a) having low affinity for β-lactam antibiotics. mecA gene is located on a mobile genetic element called staphylococcal cassette chromosome mec (SCCmec). SCCmec genomic island comprises two site-specific recombinase genes namely ccrA and ccrB [cassette chromosome recombinase] accountable for mobility. Currently, SCCmec elements are classified into types I, II, III, IV and V based on the nature of the mec and ccr gene complexes and are further classified into subtypes according to variances in their J region DNA. SSCmec type IV has been found in community-acquired isolates with various genetic backgrounds. The present study was undertaken to categorize the types of SCCmec types and subtypes I, II, III, IVa, b, c, d, and V and PVL genes among clinical MRSA isolates from COVID-19 confirmed cases.
Materials and Methods: Based on the Microbiological and Molecular (mecA gene PCR amplification) confirmation of MRSA isolated from 500 MRSA SCCmec clinical samples, 144 cultures were selected for multiplex analysis. The multiplex PCR method developed by Zhang et al. was adapted with some experimental alterations to determine the specific type of these isolates.
Results: Of the total 500 MRSA, 144 MRSA (60 were CA-MRSA and 84 were HA-MRSA) were selected for characterization of novel multiplex PCR assay for SSCmec Types I to V in MRSA. Molecular characterization of multiplex PCR analysis revealed results compare to the phenotypic results. Of the 60 CA-MRSA; in 56 MRSA strains type IVa was found and significantly defined as CA-MRSA while 4 strains showed mixed gens subtypes. Type II, III, IA, and V were present in overall 84 HA-MRSA. Molecular subtyping was significantly correlated to define molecularly as CA-MRSA and HA-MRSA however 15 (10%) strains showed mixed genes which indicates the alarming finding of changing epidemiology of CA-MRSA and HA-MRSA as well.
Conclusion: We have all witnessed of COVID-19 pandemic, and its mortality was mostly associated with co-morbid conditions and secondary infections of MDR pathogens. Rapid detections of causative agents of these superbugs with their changing epidemiology by investing in typing and subtyping clones are obligatory. We have described an assay designed for targeting SSCmec types and subtypes I, II, III, IVa,V according to the current updated SCCmec typing system. Changing patterns of molecular epidemiology has been observed by this newly described assay.
2. Chongtrakool P, Ito T, Ma XX, Kondo Y, Trakulsomboon S, Tiensasitorn C, et al. Staphylococcal Cassette Chromosome mec (SCCmec) typing of methicillin-resistant Staphylococcus aureus strains isolated in 11 Asian countries: a Proposal for a New Nomenclature for SCCmec elements. Antimicrob Agents Chemother 2006; 50: 1001-1012.
3. Strommenger B, Kehrenberg C, Kettlitz C, Cuny C, Verspohl J, Witte W, et al. Molecular characterization of methicillin-resistant Staphylococcus aureus strains from pet animals and their relationship to human isolates. J Antimicrob Chemother 2006; 57: 461-465.
4. David MZ, Daum RS. Community-associated methicillin-resistant Staphylococcus aureus: Epidemiology and clinical consequences of an emerging epidemic. Clin Microbiol Rev 2010; 23: 616-687.
5. Peterson JC, Durkee H, Miler D, Maestre-Mesa J, Arboleda A, Aguilar MC, et. al. Molecular epidemiology and resistance profile among healthcare- and community associated Staphylococcus aureus keratitis isolates. Infect Drug Resist 2019;12:831-843.
6. Yang X, Zhao J, Wang Y, Wu J, Wang X, Wang Y, et al. Molecular epidemiology of methicillin-resistant Staphylococcus aureus in Hospitalized patients in Eastern Heilongjiang Province, China. Infect Drug
Resist 2021; 14: 1635-1643.
7. Sarkhoo E, Udo EE, Boswihi SS, Monecke S, Mueller E, Ehricht R. The dissemination and molecular characterization of clonal complex 361 (CC361) methicillin-resistant Staphylococcus aureus (MRSA) in Kuwait Hospitals. Front Microbiol 2021; 12: 658772.
8. Zhang K, McClure J-A, Elsayed S, Louie T, Conly JM. Novel multiplex PCR assay for characterization and concomitant subtyping of Staphylococcal Cassette chromosome mec Types I to V in methicillin-resistant Staphylococcus aureus. J Clin Microbiol 2005; 43: 5026-5033.
9. Oliveira DC, de Lencastre H. Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Chemother 2002; 46: 2155-2161.
10. Hisata K, Kuwahara-Arai K, Yamanoto M, Ito T, Nakatomi Y, Cui L, et al. Dissemination of methicillin-resistant Staphylococci among healthy Japanese Children. J Clin Microbiol 2005; 43: 3364-3372.
11. Jadhav SV, Desai DS, Mirza SB, Apte-Deshpande AD, Deshpande S. Molecular characterization of Staphylococcal cassette chromosome mecA and concomitant Panton-valentine Leukocidine in clinical isolates of communityacquired methicillin-resistant Staphylococcus aureus. J Krishna Inst Med Sci Univ 2021; 10: 85-99.
12. Vivek JS, Nageswari Rajesh G, Mukesh S, Manpreet K, Misra RN, Matnani GB, et al. Prevalence of inducible Clindamycin resistance among community-and hospital-associated Staphylococcus aureus isolates in a tertiary care hospital in India. Biomed Res 2011; 22: 465-469.
13. Desta K, Aklillu E, Gebrehiwot Y, Enquselassie F, Cantillon D, Al-Hassan L, et al. High levels of methicillin-resistant Staphylococcus aureus carriage among healthcare workers at a teaching Hospital in Addis Ababa Ethiopia: first evidence using mecA detection. Infect Drug Resist 2022; 15: 3135-3147.
14. Monecke S, Slickers P, Gawlik D, Müller E, Reissig A, Ruppelt-Lorz A, et al. Molecular typing of ST239-MRSA-III from dverse geographic locations and the evolution of the SCCmec III element during its intercontinental spread. Front Microbiol 2018; 9: 1436.
15. Amelia S, Wahyuni DD, Yunita R, Rozi MF. The Active Surveillance of Staphylococcus aureus using Polymerase chain reaction-based identification method among Hospitalized-patient of Haji Adam Malik General Hospital, Medan, Indonesia. Open Access Maced J Med Sci 2021; 9: 622-625.
16. Mirza S, Jadhav S, Misra RN, Das NK. Coexistence of β-Lactamases in community-acquired infections in a tertiary care Hospital in India. Int J Microbiol 2019; 2019: 7019578.
17. Senok A, Somily AM, Nassar R, Garaween G, Kim Sing G, Müller E, et al. Emergence of novel methicillin-resistant Staphylococcus aureus strains in a tertiary care facility in Riyadh, Saudi Arabia. Infect Drug Resist 2019; 12: 2739-2746.
18. Mao P, Peng P, Liu Z, Xue Z, Yao C. Risk factors and clinical outcomes of hospital-acquired MRSA infections in Chongqing, China. Infect Drug Resist 2019; 12: 3709-3717.
19. Chipolombwe J, Török ME, Mbelle N, Nyasulu P. Methicillin-resistant Staphylococcus aureus multiple sites surveillance: a systemic review of the literature. Infect Drug Resist 2016; 9: 35-42.
20. Yang X, Zhao J, Wang Y, Wu J, Wang X, Wang Y, et al. Molecular epidemiology of methicillin-resistant Staphylococcus aureus in hospitalized patients in Eastern Heilongjiang Province, China. Infect Drug Resist 2021;14:1635-1643.
21. Rakonjac B, Lepšanović Z, Šuljagić V, Jovčić B, Kojić M, Larsen AR, et al. Predominance of t355/ST152/SCCmec V clonal type among PVL-positive MRSA isolates in a tertiary care hospital in Belgrade, Serbia. PLoS One 2022; 17(9): e0273474.
22. Seki M, Takahashi H, Yamamoto N, Hamaguchi S, Ojima M, Hirose T, et al. Polymerase chain reaction-based active surveillance of MRSA in emergency department patients. Infect Drug Resist 2015; 8: 113-118.
23. Tsai C-Y, Lee C-H, Chien C-C, Chen I-L. Impact of teicoplanin maintenance dose and MIC values on the clinical outcomes of patients treated for methicillin-resistant Staphylococcus aureus bacteremia. Infect Drug Resist 2018; 11: 1205-1217.
24. Udo EE, Boswihi SS, Mathew B, Noronha B, Verghese T, Al-Jemaz A, et al. Emergence of methicillin-resistant Staphylococcus aureus belonging to clonal complex 15 (CC15-MRSA) in Kuwait Hospitals. Infect Drug Resist 2020; 13: 617-626.
25. Rahi A, Kazemeini H, Jafariaskari S, Seif A, Hosseini S, Safarpoor Dehkordi F. Genotypic and phenotypic-based assessment of antibiotic resistance and profile of Staphylococcal cassette chromosome mec in the methicillin-resistant Staphylococcus aureus recovered from raw milk. Infect Drug Resist 2020; 13: 273-283.
26. Jadhav S, Jadhav V, Angadi K. Berberine: Best alternative medicine insight abating global challenges for treatment of MRSA infections [Letter]. Infect Drug Resist 2022; 15: 2573-2574.
27. Gupta N, Angadi K, Jadhav S. Molecular characterization of carbapenem-resistant Acinetobacter baumannii with special reference to carbapenemases: A systematic review. Infect Drug Resist 2022; 15: 7631-7650.
Files | ||
Issue | Vol 15 No 4 (2023) | |
Section | Original Article(s) | |
DOI | https://doi.org/10.18502/ijm.v15i4.13502 | |
Keywords | ||
Methicillin-resistant Staphylococcus aureus; SSCmec types; Staphylococcal cassette chromosome mec (SCCmec) |
Rights and permissions | |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |