BACTERIOLOGICAL AND PCR DETECTION OF PVL, MSRA AND MECA GENES AMONG STAPHYLOCOCCUS AUREUS ISOLATED FROM BURN WOUNDS

Authors

  • Shno Jalal Muhamad Surgical Teaching Hospital, Sulaimani Directorate of Health, Kurdistan Region, Iraq.
  • Khanda Abdulateef Anwar Department of Microbiology, College of Medicine, University of Sulaimani, Kurdistan Region, Iraq.
  • Sherko Ali Omer Department of Microbiology, College of Medicine, University of Sulaimani, Kurdistan Region, Iraq.

DOI:

https://doi.org/10.17656/jsmc.10230

Keywords:

Burn wound, Methicillin-resistant Staphylococcus aureus, Panton-Valentine-Leukocidin, Methionine Sulfoxide Reductase

Abstract

Background 

Methicillin-resistant Staphylococcus aureus is a pathogen that is associated with nosocomial and community- burn wound infection. S aureus produces Panton-Valentine -Leukocidin which results in the destruction of leukocytes. Resistance of S. aureus to macrolides, lincosamides, and streptogramin B is associated with the presence of an efflux pump, encoded by Methionine Sulfoxide Reductase A (msrA or msrB) genes. 

Objectives

To isolate, determine the antibiotic susceptibility pattern, and to detect the presence of pvl and msrA genes from Staphylococcus aureus, isolated from burn wounds.

Materials and Methods

A total of 423 burn wound samples (218 from hospitalized and 205 from outpatients) were cultivated on different bacteriological media. Isolates were identified and S. aureus were further subjected to antibiotic susceptibility testing using disk diffusion method. Susceptibility to methicillin, oxacillin or cefoxitin, were used to determine methicillin-resistant Staphylococcus aureus strains. Polymerase chain reaction was used to detect mecA, pvl, and msrA genes in S. aureus isolates.

Results

Bacterial growth was detected from 170 (77.9%) of hospital samples and from 183 (89.26%) community-burn wounds. The predominant isolates were Gram-negative bacilli (71.76%) among hospitalized patients followed by S. aureus (22.35%). From the community samples, Staphylococcus epidermidis was the predominant isolate (86.9%), while few species of other Gram-positive organisms were also detected but no Gram-negatives were isolated. Among the 41 S. aureus isolates, the prevalence of methicillin-resistant S. aureus strains determined by oxacillin disk diffusion method was 58.53%, 65.85% by cefoxitin, whereas, 87.8% were positive for mecA gene by PCR. Pvl was detected in 3 (7.31%), while mrsA gene was detected among 17 (41.46%) of S. aureus isolates. 

Conclusions

Infection with Methicillin-resistant Staphylococcus aureus was common in burn wounds. The prevalence of msrA gene among nosocomial and community-burn wound isolates of S. aureus was high, while few S. aureus isolates were found to carry pvl gene.

References

Plichta JK, Droho S, Curtis BJ, Patel P, Gamelli RL, Radek KA. Local burn injury impairs epithelial permeability and antimicrobial peptide barrier function in distal unburned skin. Critical Care Medicine. 2014;42(6):e420-31.2. Rafla K, Tredget EE. Infection control in the burn unit. Burns. 2011;37(1):5-15. DOI: https://doi.org/10.1097/CCM.0000000000000309

Bayram Y, Parlak M, Aypak C, Bayram I. Three-year review of bacteriological profile and antibiogram of burn wound isolates in Van, Turkey. Int J Med Sci. 2013;10(1):19-23. DOI: https://doi.org/10.7150/ijms.4723

Coban YK. Infection control in severely burned patients. World J Crit Care Med. 2012;1(4):94-101. DOI: https://doi.org/10.5492/wjccm.v1.i4.94

D'Avignon LC, Saffle JR, Chung KK, Cancio LC. Prevention and management of infections associated with burns in the combat casualty. The Journal of trauma. 2008;64(3 Suppl):S277-86. DOI: https://doi.org/10.1097/TA.0b013e318163c3e4

Dai T, Huang YY, Sharma SK, Hashmi JT, Kurup DB, Hamblin MR. Topical antimicrobials for burn wound infections. Recent Pat Antiinfect Drug Discov. 2010;5(2):124-51. DOI: https://doi.org/10.2174/157489110791233522

Uwaezuoke JC, Aririatu LE. A Survey of Antibiotic Resistant Staphylococcus Aureus Strains from Clinical Sources in Owerri. Journal of Applied Sciences and Environmental Management. 2004;8:67-9. DOI: https://doi.org/10.4314/jasem.v8i1.17230

Cui F, Li G, Huang J, Zhang J, Lu M, Lu W, et al. Development of chitosan-collagen hydrogel incorporated with lysostaphin (CCHL) burn dressing with anti-methicillin-resistant Staphylococcus aureus and promotion wound healing properties. Drug Deliv. 2011;18(3):173-80. DOI: https://doi.org/10.3109/10717544.2010.509363

CLSI. Clinical and Laboratory Standards Institute Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated From Animals; Second Informational Supplement. Clinical and Laboratory Standards Institute, Wayne, PA, USA, ISBN: 1-56238-877-0 CLSI document VET01-S2. 2013.

Lindsay JA. For CA-MRSA, How much PVL is too much? Microbiology. 2009;155(Pt 11):3473-4. DOI: https://doi.org/10.1099/mic.0.034041-0

Hu Q, Cheng H, Yuan W, Zeng F, Shang W, Tang D, et al. Panton-Valentine leukocidin (PVL)-positive health care-associated methicillin-resistant Staphylococcus aureus isolates are associated with skin and soft tissue infections and colonized mainly by infective PVL-encoding bacteriophages. Journal of Clinical Microbiology. 2015;53(1):67-72. DOI: https://doi.org/10.1128/JCM.01722-14

Adler A, Temper V, Block CS, Abramson N, Moses AE. Panton-Valentine leukocidin-producing Staphylococcus aureus. Emerging Infectious Diseases. 2006;12(11):1789-90. DOI: https://doi.org/10.3201/eid1211.060726

Tappe D, Schulze MH, Oesterlein A, Turnwald D, Muller A, Vogel U, et al. Panton-Valentine leukocidin-positive Staphylococcus aureus infections in returning travelers. American Journal of Tropical Medicine and Hygiene. 2010;83(4):748-50. DOI: https://doi.org/10.4269/ajtmh.2010.10-0252

Monecke S, Muller E, Buechler J, Rejman J, Stieber B, Akpaka PE, et al. Rapid detection of Panton-Valentine leukocidin in Staphylococcus aureus cultures by use of a lateral flow assay based on monoclonal antibodies. Journal of Clinical Microbiology. 2013;51(2):487-95. DOI: https://doi.org/10.1128/JCM.02285-12

Lim JC, You Z, Kim G, Levine RL. Methionine sulfoxide reductase A is a stereospecific methionine oxidase. Proc Natl Acad Sci U S A. 2011;108(26):10472-7. DOI: https://doi.org/10.1073/pnas.1101275108

Juda M, Chudzik-Rzad B, Malm A. The prevalence of genotypes that determine resistance to macrolides, lincosamides, and streptogramins B compared with spiramycin susceptibility among erythromycin-resistant Staphylococcus epidermidis. Memorias do Instituto Oswaldo Cruz. 2016;111(3):155-60. DOI: https://doi.org/10.1590/0074-02760150356

Connie RM, donald CL, George M. Textbook of Diagnostic Microbiology 3rd ed: Newark Saunders Elsevier; 2007. p. 303-15.

Sabzghabaee AM, Abedi D, Fazeli H, Javadi A, Jalali M, Maracy MR, et al. Antimicrobial resistance pattern of bacterial isolates from burn wounds in an Iranian University Hospital. J Res Pharm Pract. 2012;1(1):30-3. DOI: https://doi.org/10.4103/2279-042X.99675

Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 1966;45(4):493-6. DOI: https://doi.org/10.1093/ajcp/45.4_ts.493

Anyanwu NCJ, Abdullahi IO, Ameh JB, Ella EE. Molecular detection of PVL, msrA genes and antibiotic susceptibility pattern of staphylococcus aureus from skin and soft tissue infections in Zaria, Nigeria. Scientific Journal of Microbiology. 2013;2(2):43-52.

Elhassan MM, Ozbak HA, Hemeg HA, Elmekki MA, Ahmed LM. Absence of the mecA Gene in Methicillin Resistant Staphylococcus aureus Isolated from Different Clinical Specimens in Shendi City, Sudan. Biomed Res Int. 2015;2015:895860. DOI: https://doi.org/10.1155/2015/895860

Lee PY, Costumbrado J, Hsu CY, Kim YH. Agarose gel electrophoresis for the separation of DNA fragments. J Vis Exp. 2012(62). DOI: https://doi.org/10.3791/3923

Jebur M. Therapeutic efficacy of Lactobacillus acidophilus against bacterial isolates from burn wounds. N Am J Med Sci. 2010;2(12):586-91. DOI: https://doi.org/10.4297/najms.2010.2586

Hussien IA, Habib KA, Jassim KA. Bacterial Colonization of Burn Wounds. Baghdad Science Journal. 2012;9(4). DOI: https://doi.org/10.21123/bsj.9.4.623-631

Shrivastava G, Bhatambare GS, Lunawat A, Patel KB, Maheshwari T. Pattern of microorganism from burn wounds and their trends of susceptibility to antibiotics in tertiary care center. Int J Health Syst Disaster Manage. 2016;4(2):53-7. DOI: https://doi.org/10.4103/2347-9019.183229

Mohamed H. One year prevalence of critically ill burn wound bacterial infections in surgical ICU in Egypt: Retrospective study. Egyptian Journal of Anaesthesia. 2016;32(3):431–4. DOI: https://doi.org/10.1016/j.egja.2016.01.005

Irfan M, Ahmed I, Shafee M, Tareen AM, Rehman MFU, Khan SA. Microbiological Investigation of Burn Patients in Burn Intensive Units, in Quetta, Pakistan. Journal of Infection and Molecular Biology. 2014;2(4):74 – 6. DOI: https://doi.org/10.14737/jimb.2307-5465/2.4.74.76

Kareem SM, Al- Jubori SS, Ali MR. Prevalence of PVL Gene among Methicillin Resistance S. aureus Isolates in Baghdad City. World Journal of Pharmaceutical Research. 2015;4(5):455-71.

Ahmad B, Khan F, Ahmed J, Cha SB, Shin M, Bashir S, et al. Antibiotic Resistance Pattern and Molecular Epidemiology of Methicillin-Resistant Staphylococcus aureus Colonization in Burns Unit of a Tertiary Care Hospital in Peshawar, Pakistan. Tropical Journal of Pharmaceutical Research. 2014;13(12):2091-9. DOI: https://doi.org/10.4314/tjpr.v13i12.21

Ragbetli C, Parlak M, Bayram Y, Guducuoglu H, Ceylan N. Evaluation of Antimicrobial Resistance in Staphylococcus aureus Isolates by Years. Interdiscip Perspect Infect Dis. 2016;2016:9171395. DOI: https://doi.org/10.1155/2016/9171395

Degaim ZD, Shani WS, Hamim SS. Virulence factors of Methicillin Resistant Staphylococcus aureus (MRSA) isolated from burn patients. IntJCurrMicrobiol. 2015;4(7):898-906.

Ohadian Moghadam S, Pourmand MR, Aminharati F. Biofilm formation and antimicrobial resistance in methicillin-resistant Staphylococcus aureus isolated from burn patients, Iran. J Infect Dev Ctries. 2014;8(12):1511-7. DOI: https://doi.org/10.3855/jidc.5514

Kolhe VS, R. M, M. MS. Incidence of Antibiotic Resistance in Methicillin Resistant Staphylococcus aureus In Burn Patients in a Tertiary Care Hospital, India. International Journal of Pharma and Bio Sciences. 2014;5(4):537 - 41.

Pramodhini S, Thenmozhivalli PR, R. S, Dillirani V, Vasumathi A, Agatha D. Comparison of Various Phenotypic Methods and mecA Based PCR for the Detection of MRSA. Journal of Clinical and Diagnostic Research. 2011;5(7):1359 - 62.

Demir T, Coplu N, Esen B. Comparative analysis of phenotypic and genotypic detection of methicillin resistance among Staphylococcus aureus. Indian Journal of Pathology and Microbiology. 2016;59(3):314-7. DOI: https://doi.org/10.4103/0377-4929.188103

Broekema NM, Van TT, Monson TA, Marshall SA, Warshauer DM. Comparison of cefoxitin and oxacillin disk diffusion methods for detection of mecA-mediated resistance in Staphylococcus aureus in a large-scale study. Journal of Clinical Microbiology. 2009;47(1):217-9. DOI: https://doi.org/10.1128/JCM.01506-08

Farahani A, Mohajeri P, Gholamine B, Rezaei M, Abbasi H. Comparison of different phenotypic and genotypic methods for the detection of methicillin-resistant Staphylococcus aureus. N Am J Med Sci. 2013;5(11):637-40. DOI: https://doi.org/10.4103/1947-2714.122305

Khosravi AD, Hoveizavi H, Farshadzadeh Z. The prevalence of genes encoding leukocidins in Staphylococcus aureus strains resistant and sensitive to methicillin isolated from burn patients in Taleghani Hospital, Ahvaz, Iran. Burns. 2012;38(2):247-51. DOI: https://doi.org/10.1016/j.burns.2011.08.002

Qureshi A, Ingle R, Musaddiq M, Ali Y, Khan Z. Status and Distribution of mecA Gene in Hospitalized Patient’s MRSA Isolates. Bioscience Discovery. 2012;3:52-7.

DeLeo FR, Chambers HF. Reemergence of antibiotic-resistant Staphylococcus aureus in the genomics era. Journal of Clinical Investigation. 2009;119(9):2464-74. DOI: https://doi.org/10.1172/JCI38226

Shariati L, Validi M, Hasheminia AM, Ghasemikhah R, Kianpour F, Karimi A, et al. Staphylococcus aureus Isolates Carrying Panton-Valentine Leucocidin Genes: Their Frequency, Antimicrobial Patterns, and Association With Infectious Disease in Shahrekord City, Southwest Iran. Jundishapur J Microbiol. 2016;9(1):e28291. DOI: https://doi.org/10.5812/jjm.28291

AL-Charrakh AH, AL-Hassnawi HH, AL-Khafaji JK. Molecular Characteristics of Community-Associated Methicillin-resistant Staphylococcus aureus (CAMRSA) Isolates from Clinical Specimens in Iraq. British Microbiology Research Journal. 2015;5(3):227-36. DOI: https://doi.org/10.9734/BMRJ/2015/9607

Zmantar T, Kouidhi B, Miladi H, Bakhrouf A. Detection of macrolide and disinfectant resistance genes in clinical Staphylococcus aureus and coagulase-negative staphylococci. BMC Res Notes. 2011;4:453. DOI: https://doi.org/10.1186/1756-0500-4-453

Fasihi Y, Saffari F, Ghahraman MRK, Neyestanaki DK. Molecular Detection of Macrolide and Lincosamide-Resistance Genes in Clinical Methicillin-Resistant Staphylococcus aureus Isolates from Kerman, Iran. Arch Pediatr Infect Dis. 2017;5(1):1-5. DOI: https://doi.org/10.5812/pedinfect.37761

Published

2019-12-21

How to Cite

1.
Muhamad S, Anwar K, Omer S. BACTERIOLOGICAL AND PCR DETECTION OF PVL, MSRA AND MECA GENES AMONG STAPHYLOCOCCUS AUREUS ISOLATED FROM BURN WOUNDS. JSMC [Internet]. 2019 Dec. 21 [cited 2024 May 27];9(4):391-400. Available from: https://jsmc.univsul.edu.iq/index.php/jsmc/article/view/jsmc-10230

Similar Articles

1-10 of 36

You may also start an advanced similarity search for this article.

Most read articles by the same author(s)

1 2 > >>