Molecular detection of mecA (or mecC) gene in MR-CoNS isolated from infected patients in Hospital Tengku Ampuan Afzan and International Islamic University Malaysia Medical Centre

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Research Paper 17/04/2023
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Molecular detection of mecA (or mecC) gene in MR-CoNS isolated from infected patients in Hospital Tengku Ampuan Afzan and International Islamic University Malaysia Medical Centre

Abdillah Mussa Kitota
Int. J. Biomol. & Biomed.16( 2), 1-19, April 2023.
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The emergence of nosocomial infections caused by CoNS has led clinicians and researchers to reconsider the role of CoNS and methicillin-resistant CoNS (MR-CoNS) as important agents of nosocomial infections. The present study was conducted on clinical isolates of MR-CoNS obtained from inpatients in Hospital Tengku Ampuan Afzan (HTAA) and International Islamic University Malaysia Medical Center (IIUM-MC) to detect the presence of mecA or mecA homologue (mecC gene). A total of 40 isolates (33 blood, 4 tissues, and 3 swabs) of MR-CoNS were collected through venepuncture, biopsy, and swabbing techniques respectively, and processed by conventional cultural and biochemical methods, antimicrobial susceptibility tests, and finally confirmation to the species level was done by using conventional PCR assay for known four common clinical species. Methicillin and vancomycin-resistance profile of the isolates was performed by E-test and broth micro-dilution methods. Of the 40 isolates, 38 were identified to be methicillin-resistant (MIC ³ 0.5µg/mL). The remaining 2 isolates were considered as susceptible to methicillin (MIC ≤ 0.25). All 40 isolates were found to be susceptible to vancomycin, with MIC ranging from 1-4µg/mL. Only mecA was identified in the 38 MR-CoNS isolates (95%). The other 2 isolates (5%) that were identified to be methicillin-sensitive by the E-test, also tested negative for the presence of mecC gene, thus confirmed to be non-methicillin resistant. The high percentage of mecA gene among these MR-CoNS isolates points toward the need for periodic antibiogram surveillance as they are identified to cause difficult to treat infections.


Akpaka PE, Smikle MF. 2007. Molecular Epidemiology of Methicillin Resistant Coagulase-negative Staphylococci Isolates from Blood Specimens at a Tertiary Hospital in the Caribbean. West Indian Medical Journal 56(5), 464.

Al Tayyar IA, Al-Zoubi MS, Hussein E, Khudairat S, Sarosiekf K. 2015. Prevalence and antimicrobial susceptibility pattern of coagulase-negative staphylococci (CoNS) isolated from clinical specimens in Northern of Jordan. Iranian Journal of Microbiology 6(7), 294-301.

Albertsen M, Brandt J. 2018. Investigation of Detection Limits and the Influence of DNA Extraction and Primer Choice on the Observed Microbial Communities in Drinking Water Samples Using 16S rRNA Gene Amplicon Sequencing. Frontier in microbiology 9, 2140.

Alicem T, Tuba D, Ozcan D, Recap T, Nida, O, Selahattin A, Saim D. 2014. In vitro susceptibility to methicillin, vancomycin and linezolid of staphylococci isolated from bloodstream infections in eastern Turkey. Brazilian Journal of Microbiology 45(3), 829-833.

Argemi X, Hansmann Y, Prola K, Prevost G. 2019. Coagulase-Negative Staphylococcus Pathogenomics. International Journal of Molecular Sciences 20, 1215.

Baird-Parker AC. 1965. The Classification of Staphylococci and Micrococci from World-Wide Sources. Journal of General Microbiology 38, 363-87.

Bannerman TL. 2003. Staphylococcus, Micrococcus, and other catalase-positive cocci that grow aerobically. In Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH (eds), Manual of Clinical Microbiology, American Society Microbiology, Washington 384-404.

Beck WD, Berger-Bacchi, Kayeser FH. 1986. Additional DNA in methicillin-resistant Staphylococcus aureus and Molecular cloning of mec-specific DNA. Journal of bacteriology 165, 373-378.

Becker K, Heilmann C, Peters G. 2014. Coagulase-Negative Staphylococci. Clinical Microbiology Review 27(4), 870-926. https:// PMID: 25278577.

Begum ES, Anbumani N, Kalyani J, Mallika M. 2011. Prevalence and antimicrobial susceptibility pattern of Coagulase-negative Staphylococcus. International journal of medical pharmacology 1(4), DOI: 10.5530/ijmedph.4.2011.14.

Brook GF, Carrol KC, Butel JS., Morse SA, Mietzner TA. 2010. Jewertz, Menelick & Adelberg’s Medical Microbiology. McGrewHill Companies. International Edition. 25 ed. pp. 185.

Bugum S, Anbumani N, Kalyani J, Mallika J. 2011. Prevalence and antimicrobial susceptibility pattern of Coagulase-negative Staphylococcus. International Journal of Medicine and Public health 1(4), 59-62.

Cheung GYC, Otto M. 2010. Understanding the significance of Staphylococcus epidermidis bacteraemia in babies and children. Current Opinion of Infectious Diseases 23, 208-216.

Christensen GD, Simpson WA, Bisno AL, Beachey EH. 1983. Adherence of slime-producing strains of Staphylococcus epidermidis to smooth surfaces. Infectious Immunology 37, 318-326.

Christensen GD. 1993. The sticky problem of Staphylococcus epidermidis. Sepsis Hospital Practice 30, 27-38.

Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing. 16th Informational Supplement. CLSI document M100, 28th ed. Wayne, PA 19087 USA. Clinical Infectious Disease 32, S114-S132. [PubMed: 11320452].

Cucarella C, Tormo MA, Ubeda C, Trotonda MP, Monzon M, Peris C. 2004. Role of biofilm-associated protein Bap in the pathogenesis of bovine Staphylococcus aureus. Infectious Immunology 72(4), 2177-2185. 4.2177-2185.2004 PMID: 15039341.

Cunha SR, LM, Sinzato KY, Liciana, VA, Silveira AV. 2004. Cunha Comparison of Methods for the Identification of Coagulase-negative Staphylococci. Memories do Instituto Oswaldo Cruz, Rio de Janeiro 99(8), 855-860.

Cunha SRLM, Rugolo SSML, Lopes AC. 2006. Study of virulence factors in coagulase-negative staphylococci isolated from newborns. Memories do Instituto Oswaldo Cruz, Rio de Janeiro 101(6), 661-668.

D’mello D, Andrew J, Rahman SM, Qu Y, Garland S, Pearce C, Deighton AM. 2008. Vancomycin Heteroresistance in Bloodstream Isolates of Staphylococcus capitis. Journal of Clinical Microbiology 46(9), 3124-3126.

Deyno S, Fekadu S, Seyfe S. 2018. Prevalence and antimicrobial resistance of coagulase negative staphylococci clinical isolates from Ethiopia: a meta-analysis. BMC Microbiology 18, 43

Diekema DJ. 2001. Survey of infections due to Staphylococcus species: frequency of occurrence and antimicrobial susceptibility of isolates collected in the United States, Canada, Latin America, Europe, and the Western Pacific region for the SENTRY Antimicrobial Surveillance Program 1997-1999.

Diep BA. 2006. Complete genome sequence of USA300, an epidemic clone of community-acquired methicillin-resistant Staphylococcus aureus. Lancet 367, 731-739. [PubMed: 16517273].

Du X, Zhu Y, Song Y, Li T, Luo T. 2013. Molecular Analysis of Staphylococcus epidermidis Strains Isolated from Community and Hospital Environments in China. PLoS ONE 8(5), e62742.

Dupont CV, Sivadon TEB, Ille B, Dauphin, Beretti JL. 2010. Identification of clinical coagulase negative Staphylococci, isolated in microbiology laboratories, by matrix assisted laser desorption/ionization time of flight mass spectrometry and two automated systems. Clinical Microbiology Infections 16, 998-1004.

Espy MJ, Uhl JR, Sloan LM, Bulkwalter SP, Jones MF, Volter EA, Yao JD, Wangenack NL, Rosenblatt JE, Cockerill FR, Smith TF. 2006. Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clinical Microbiology Review 19(3), 595.

Foster T. 1996. Medical Microbiology, 4th edition.

Garrett DO, Jochimsena E, Murfitt K, Hill B, McAllister S, Nelson P, Spera PV, Sall RK, Tenover FC, Johnston J, Zimmer B, Jarvis WR. 1999. The emergence of decreased susceptibility to vancomycin in Staphylococcus epidermidis. Infection Control and Hospital Epidemiology 20(3), 167-70.

Gilani M, Usman J, Latif M, Munir T, Gill MM, Anjum R, Babar N. 2016. Methicillin resistant coagulase negative Staphylococcus: From colonizer to a pathogen. Pakistan Journal of Pharmaceutical Sciences 26(4), 1117-1121.

Grace JA, Olayinka BO, Onaolapo JA, Obaro SK. 2019. Staphylococcus aureus and Coagulase-Negative Staphylococci in Bacteraemia: The Epidemiology, Predisposing Factors, Pathogenicity and Antimicrobial Resistance. Clinical Microbiology: Open access 8, 325. DOI: 10.4172/2327

Gras-Le GC, Fournier S, Andre-Richet B. 2007. Almond oil implicated in a Staphylococcus capitis outbreak in a neonatal intensive care unit. Journal of Perinatology 27(11), 713-7.

Guyer RL, Koshland DE. 1989. The Molecule of the year. Journal of science 246, 1543-1546.

Hanssen AM, Kjeldsen G, Sollid JU. 2004. Local variants of Staphylococcal cassette chromosome mec in sporadic methicillin-resistant Staphylococcus aureus and methicillin-resistant coagulase-negative Staphylococci: evidence of horizontal gene transfer? Antimicrobial Agents Chemotherapy 48, 285-96.

Hedman P, Ringertz O, Eriksson B, Kvarnfors P, Andersson M, Bengtsson L, Olsson K. 1990. Staphylococcus saprophyticus found to be a common contaminant of food. Journal of Infectious disease 21, 11-19. .1016/0163-4453(90)

Heilmann C, Ziebuhr W, Becker K. 2018. Are Coagulase-negative staphylococci virulent? Clinical Microbial Infections 25(9), 1071-1080. DOI: 10.1016/j.cmi.2018.11.012.

Herman-Bausier P, El-Kirat-Chatel S, Foster TJ, Geoghegan JA, Dufrêne YF. 2015. Staphylococcus aureus Fibronectin-Binding Protein A mediates cell-cell adhesion through low-affinity homophilic bonds. Molecular Biology 6(3), e00413-15. PMID:

Huebner H, Goldmann AD. 1999. Coagulase negative Staphylococci: Role as Pathogens. Annual Review of Medicine 50, 223.

Jain N, Mathur P, Misracm. 2013. Trend of vancomycin susceptibility of staphylococci at a level 1 trauma centre of India. Indian Journal of Medical Research 136(6), 1022-1024.

James H, Jorgensen, Mary Jane Ferraro. 2009. Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clinical infectious disease 49, 1749-55.

Jan-Roblero J, Rodriguez-Martínez S, Cancino-Diaz EM, Juan C, Cancino-Diaz CJ. 2016. Staphylococcus Biofilms Licensee InTech.

Jin X, Yue S, Wells KS, Singer VL. 1994. SYBR Green: I. A new fluorescent dye optomized for detection of picogram amounts of DNA in gels. Biophysical Journal 66, A159.

Jorgensen JH, Ferraro MJ. 2009. Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clinical Infectious Diseases 49(11), 1749-1755.

Klingenberg C, Ronnestad A, Anderson AS. 2007. Persistent strains of coagulase-negative staphylococci in a neonatal intensive care unit: virulence factors and invasiveness. Clinical Microbiology Infections 13, 1100-11.

Kloos WE, Bannerman LT. 1994. Updates on Clinical Significance of Coagulase-Negative Staphylococci. Clinical Microbiology Reviews 117-140.

Kloos WE, Schleifer KH. 1975. Simplified scheme for routine identification of human Staphylococcus species. Journal of Clinical Microbiology 1(1):82-8.

Kloos WE. 1980. Natural populations of the genus Staphylococcus. Annual Review of Microbiology 34, 559-92.

Koksal F, Yasar H, Samasti M. 2009. Antibiotic resistance patterns of coagulase-negative staphylococcus strains isolated from blood cultures of septicemic patients in Turkey. Microbiology Research 164, 404-10.

Kroos WE, Bennerman TL. 1994. Update on clinical significance of coagulase-negative staphylococci. Clinical Microbiology review 7(1), 117-40.

Kryndushkin DS, Alexandrov IM, Ter-Avanesyan MD, Kushnirov VV. 2003. Yeast [PSI+] prion aggregates are formed by small Sup35 polymers fragmented by Hsp10. Journal of Biological Chemistry 278(49), 49636.

Laham NA. 2017. Species Identification of Clinical Coagulase-negative Staphylococci Isolated in Al-Shifa Hospital Gaza using Matrix-assisted Laser Desorption/Ionization-time of Flight Mass Spectrometry. Current Research Bacteriology, ISSN 1994-5426.

Larson RA, Burke JP. 1986. The epidemiology and risk factors for nosocomial catheter associated bacteriuria caused by Coagulase negative Staphylococci. Infectious Control 7, 212-215.

Lee PY, Costumbrado J, Hsu CY, Kim YH. 2012. Agarose Gel Electrophoresis for the Separation of DNA Fragments. Journal of visualized experiment 62, e3923. DOI:10.3791/3923.

Lodge J, Lund P, Minchin S. 2007. Gene cloning: principles and applications. ISBN 0- 7487-6534-4.

Longauerova A. 2006. Coagulase negative staphylococci and their participation in pathogenesis of human infections. Bratislavske Lekarske Listy 107, 448-52.

Lorenz TC. 2012. Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies. Journal of Visualized Experiments 63, e3998. DOI: 10.3791/3998.

Ma XX. 2002. Novel type of staphylococcal cassette chromosome mec identified in community- acquired methicillin-resistant Staphylococcus aureus strains. Antimicrobial Agents Chemotherapy 46, 1147-1152. [PubMed: 11897611].

Madruga MH, Moscatello DK, Emlet DR, Dıeterıch R, Wong AJ. 1997. Grb2 associated binder mediates phosphatidylinositol 3-kinase activation and the promotion of cell survival by nevre growth factor. Proceedings of the National Academy of Sciences 94, 12419-12424.

Maira-Litra ́n T, Kropec A, Abeygunawardana C, Joyce J, Mark G, Goldmann DA. 2002. Immunochemical properties of the staphylococcal poly-N-acetylglucosamine surface polysaccharide. Infection and Immunology 70(8), 4433-4440. 8.4433-4440.2002 PMID: 12117954.

Martinez-Meléndez A, Morfin-Otero R, Villarreal-Trevino L, Camacho-Ortiz A, Gonzalez-Gonzalez G, Llaca-Diaz J, Rodríguez-Noriega E, Garza-González E. 2016. Molecular epidemiology of Coagulase-negative bloodstream isolates: detection of Staphylococcus epidermidis ST2, ST7 and linezolid-resistant ST23. The Brazilian Journal of infectious disease 20(5), 419-428.

Matheson M. 1999. Gram stain. Community eye health Journal 12(30), 24 PMCID: PMC1706001. PMID: 17491986.

Mir BA, Srikanth. 2013. prevalence and antimicrobial susceptibility of methicillin resistant Staphylococcus aureus and coagulase-negative staphylococci in a tertiary care hospital. Asian Journal of Pharmaceutical and Clinical Research 6(3), SSN – 0974-2441.

Miragaia M. 2002. Molecular characterization of methicillin-resistant Staphylococcus epidermidis clones: evidence of geographic dissemination. Journal of Clinical Microbiology 40, 430-438.

Miragaia M, De Lencastre H, Perdreau-Remington F. 2009. Genetic diversity of arginine catabolic mobile element in Staphylococcus epidermidis. PloS one 64(11), e7722.

Miragaia M, Thomas JC, Couto I, Enright MC, de Lencastre H. 2007. Inferring a population structure for Staphylococcus epidermidis from multilocus sequence typing data. Journal of Bacteriology 189, 2540-2552.

Mullis K, Faloona FA. 1987. Specific synthesis of DNA in vitro via a polymerase catalyzed chain reaction. Methodology in Enzymology 255, 335-350.

Murray PR, Barron EJ, Jorgenson JH., Pfaller MA, Yolken RH. 2003. Manual of Clinical Microbiology. 8th Edition. Volume 1. U.S.A. Library of Congress-in-Publication Data.

Murray PR. 2010. Matrix assisted laser desorption ionization time of flight mass spectrometry: Usefulness for taxonomy and epidemiology. Clinical Microbiology. Infections 16, 1626-1630.

Nahaei RM, Shahmohammadi RM, Ebrahim S, Milani M. 2015. Detection of Methicillin-Resistant Coagulase Negative Staphylococci and Surveillance of Antimicrobial Resistance in a Multi-Center Study from Iran. Jundishapur Journal of Microbiology 8(8), e19945.

Natoli S, Fontana C, Favaro M, Bergamini A, Testore GP, Minelli S. 2009. Characterization of coagulase-negative staphylococcal isolates from blood with reduced susceptibility to glycopeptides and therapeutic options. Biomedical Journal of Infectious diseases 9, 83.

Osmon DR, Sampathkumar P, Cockerill FR. 2000. Prosthetic joint infection due to Staphylococcus lugdunensis. Mayo Clinic Proceedings 75, 511-512.

Otto M. 2004. Virulence factors of the coagulase negative staphylococci. Front Biosciences 9, 841-63.

Otto M. 2008. Staphylococcal biofilms. Current Topics in Microbiology and Immunology 322, 207-228. PMCID: PMC2777538 PMID: 18453278.

Otto M. 2009. Staphylococcus epidermidis – the ‘accidental’ pathogen. Nature Reviews Microbiology 7, 555-567.

Otto M. 2013. Coagulase-negative staphylococci as reservoirs of genes facilitating MRSA infection: Staphylococcal commensal species such as Staphylococcus epidermidis are being recognized as important sources of genes promoting MRSA colonization and virulence. Bioessay 35(1), 4-11. doi:10.1002/bies.201200112.

Pfaller MA, Herwaldt LA. 1988. Laboratory, Clinical, and Epidemiological As- pects of Coagulase-Negative Staphylococci. Clinical Microbial Reviews 1, 281-299.

Rahman ZA, Hamzah SA, Hassan AM, Osman S, Siti Suraiya Md Noor SSN. 2002. The significance of coagulase-negative staphylococci bacteremia in a low resource setting. Journal of Infection in Developed Countries 7(6), 448-452.

Rania AA, Nsreen MK, Rasha HEl, Mona MA. 2017. Evaluation for the Novel mecC Methicillin Resistance among Methicillin Resistant Staphylococcal Isolates in two Egyptian University Hospitals. Archieve of Clinical Microbiology 9(1), 71.

Reece R. 2004. Analysis of gene and genome. NJ 07030, USA, Willey Ltd.

Reffuveille F, Josse J, Vallecm, Gangloff CS. 2017. Staphylococcus aureus Biofilms and their Impact on the Medical Field. Open access peer review chapter. DOI: 10.5772/66380.

Ryan KJ, Ray CG (editors). 2004. Sherris Medical Microbiology. 4th edition. McGraw Hill. SBN 0-8385-8529-9.

Saber H, Jasni AS, Maztura TZ, Jamaluddin T, Ibrahim R. 2017. A Review of Staphylococcal Cassatte Chromosomae mec (SCCmec) Types in Coagulase-Negative Staphylococci (CoNS) Species. Malays Journal of Medical Sciences 24(5), 7-18.

Sani NM, Sapri HF, Noorin A, Neoh H, Hussin S. 2011. Species Identification of Coagulase Negative Staphylococci (CoNS) Isolates in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). Asia-Pacific Journal of Molecular Medicine 1, 5.

Sanzen L, Walder M. 1988. Antibiotic resistance of Coagulase negative Staphylococci in an orthopaedic department. Journal of Hospital Infections 12, 103-108.

Sarathbabu R, Rajkumari, Ramani T. 2013. Characterization of Coagulase Negative Staphylococci isolated from urine, pus, sputum and blood stream. International Journal of Pharmaceutical Science Invention 2(1), 37-46.

Schumecher A, Vranken T, Malhotra A, Arts JJC, Habibovic P. 2018. In vitro antimicrobial susceptibility testing methods: agar dilution to 3D tissue-engineered models. European Journal of Clinical Microbiology and Infectious Disease 37(2), 187-208. DOI: 10.1007/s10096-017-3089-2.

Schwarz S, Silley P, Simjee S, Woodford N, van Duijkeren E, Johnson A, Gaastra W. 2010. Editorial: Assessing the antimicrobial susceptibility of bacteria obtained from animals. Journal of Antimicrobial Chemotherapy 65, 601-604.

Seng R, Kitti T, Thumeepak R, Kongthai P, Leungtongkam U, Wannalerdsakun S, Sitthisak S. 2017a. Biofilm formation of methicillin-resistant coagulase negative staphylococci (MR-CoNS) isolated from community and hospital environments. PLoS ONE 12(8), e0184172. https://

Seng R, Leungtongkam U, Thummeepak R, Chatdumrong W, Sitthisak S. 2017b. High prevalence of methicillin-resistant coagulase-negative Staphylococci isolated from a university environment in Thailand. International Microbiology 20(2), 65-73.

Sharma V, Jindai N, Devi P. 2010. Prevalence of methicillin resistant coagulase negative staphylococci in a tertiary care hospital. Iranian Journal of Microbiology 2(4), 185-188.

Sharp PA, Sugden B, Sambrook J. 1973. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresis. Biochemistry 12, 3055-3063.

Soumya KR, Philip S, Sugathan S, Mathew J, Radhakrishnan EK. 2017. Virulence factors associated with Coagulase Negative Staphylococci isolated from human infections. 3 Biotechnology 7, 140. DOI 10.1007/s13205-017-0753-2.

Ternes YM, Juliana Lamaro-Cardoso J, André MCP, Pessoa JRVP, Maria Vieira MAS, Minamisava R, Andrade AL, Kipnis A. 2013. Molecular epidemiology of coagulase-negative Staphylococcus carriage in neonates admitted to an intensive care unit in Brazil. BMC Infectious Diseases 13, 572.

Thomas KR, Melton R, Asbell AP. 2019. Antibiotic resistance among ocular pathogens: current trends from the ARMOR surveillance study (2009-2016). Clinical Optometry 1, 15-26.

Vandenesch F, Etienne J, Reverdy ME, Eykyn SJ. 1993. Endocarditis due to the Staphylococcus lugdunensis: report of 11 cases and review. Clinical Infectious Diseases 17, 871-876.

Widerström M. 2010. Molecular epidemiology of coagulase-negative staphylococci in hospitals and in the community. Print & Media, Umeå University, Umeå, Sweden. ISBN: 978-91-7459-020-3. ISSN: 0346-6612.

Winslow W. 1908. The Systematic Relationships of the Coccaceae. New York: John Rily.

Zhou X, Li Y. 2015. Atlas of Oral Microbiology: From healthy microflora to disease 32 Jamestown Road, London: Elsevier Inc.