Molecular detection of methicillin resistant Staphylococcus aureus harbouring β- lactamase resistance genes isolated from different sources of infections

Authors

  • Amir Hani Raziq Scientific Research Center, College of Sciences, University of Duhok, Duhok, Iraq.
  • Ahmed Mohammed Sulaiman Scientific Research Center, College of Sciences, University of Duhok, Duhok, Iraq
  • Narmin Saeed Merza Scientific Research Center, College of Sciences, University of Duhok, Duhok, Iraq
  • Zainab Hasan Abdullah Scientific Research Center, College of Sciences, University of Duhok, Duhok, Iraq

DOI:

https://doi.org/10.15218/zjms.2017.048

Keywords:

Methicillin resistant Staphylococcus aureus, mec A, TEM, multiplex- PCR, Uniplex-PCR

Abstract

Background and objective: The detection and investigation of methicillin resistance staphylococci specifically S. aureus in clinical microbiology setting is very helpful both for informing the appropriate treatment of individual patients and also for the surveillance of these organisms. This study aimed at the rapid molecular detection of methicillin resistant staphylococci harbouring β-lactamase gene and determination of the efficiency of m-PCR through comparison with uniplex PCR.

Methods: Standard microbiological techniques were applied for the determination of the presence of methicillin resistant S. aureus in samples recovered from different body sites of patients who attended Al-Kadhumyhia Teaching and Baghdad Teaching Hospitals. The resulting methicillin resistant S. aureus (MRSA) isolates were subjected to uni and multiplex PCR amplifications for detecting the existence of mec A gene and β-lactamase (TEM) resistance gene.

Results: Half of the cases involved were found to be caused by MRSA. All the tested isolates showed positive amplification bands for the presence of mec A gene and only 48.8% of these harbored TEM gene. The obtained results revealed high sensitivity of universal bacterial and TEM primers expressed as 97.6% and 100% respectively, while the sensitivity of mec A primer was limited to 60%.

Conclusion: The phenotypic identification of MRSA revealed a higher incidence rate and that different molecular techniques can yield conflicting results and it can also be concluded that resistant due to beta- lactamase production can be a crucial factor added to the previously settled methicillin resistant due to mec A gene.

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References

Khadri H, Alzohairy M. Prevalence and antibiotic susceptibility pattern of methicillin-resistant and coagulase-negative staphylococci in a tertiary care hospital in India. Int J Med Med Sci 2010; 2(4):116-20.

Chambers HF. Methicillin resistance in staphylococci: Molecular and biochemical basis and clinical implications. Clin Microbiol Rev 1997; 10:781-91.

Samah SE, Mohamad S, Mohamad S. Accuracy of Phenotypic Methods in Detection of Methicillin Resistant Staphylococci Species Compared to PCR Assay for mecA Gene. Egypt J Med Micro 2009; 18(1):33-40.

Samah SE, Mohamad S, Mohamad S. Accuracy of Phenotypic Methods in Detection of Methicillin Resistant Staphylococci Species Compared to PCR Assay for mecA Gene. Egypt J Med Micro 2009; 18(1).

Matthews PR, Stewart PR. Resistance heterogeneity in methicillin-resistant Staphylococcus aureus. FEMS Microbiol Lett 1984; 22:161-6.

Ann H, Pierre L, Francois JP, Marthe B, Martin G, Nathalie B, et al. Identification of Methicillin-Resistant Staphylococcus aureus Carriage in Less than 1 Hour during a Hospital Surveillance Program. Clin Infect Dis 2005; 40:976-81.

Baron EJ, Peterson LR, Finecold SM. Diagnostic microbiology. 5 th ed. Toronto Canada: The C.V. Mesby Company; 1994. P. 429-43.

Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. Fifteenth informational supplement, document M100-S15. CLSI, Wayne, Pa, USA; 2005.

Al-Mizory S. Study of the Bacterium Pseudomonas aeruginosa Isolated from Different Sources at Dohuk City. M. Sc. Thesis. College of Education, Dohuk University. Iraq; 2007.

Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: A laboratory manual. 2nd ed. New York: Cold spring harbor laboratory press;1989.

Sadeghian S, Neyestani TR, Burnie JP. Detection of Bacterial, Methicillin resistance, and b. Lactamase enzyme gene found in wound swabs by Multiplex Polymerase Chain Reaction. Acta Medica Iranica 2004; 42(1):19-25.

Berger-Bachi, B. Resistance mechanisms of Gram positive bacteria. Int J Med Microbiol 2002; 292(1):27-35.

Petinaki E, Arvaniti A, Bartzavali C, Dimitracopoulos G, Spiliopoulou I. Presence of mec genes and overproduction of β-lactamase the expression of low-level methicillin resistance among staphylococci. Chemotherapy 2002; 48(4):174-81.

Corrente M, Greco G, Madio A, Ventrigila G. Methicillin resistance in staphylococci isolated from subclinical mastitis in sheep. New Microbiol 2003; 26(1):39-45.

Perez-Perez FJ, Hanson ND. Detection of plasmid-mediated AmpC beta-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol 2002; 40(6):2153-62.

Ribas RM, Gontijo-Filho PP, Darini LC. Conventional versus molecular tests (multiplex PCR and PCR mec A gene) for detection of methicillin resistant Staphylococcus aureus. Braz J Microbiol 2003; 34 (Suppl.1):35-7.

Martineau F, Picard FJ, Lansac N, Nenard C, Roy PH, Ovellehe M, et al. Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and S. epidermidis. Antimicrobial Agents Chemother 2000; 44:231-8.

Mohanasoundaram KM, Lalitha MK. Comparison of phenotypic versus genotypic methods in the detection of methicillin resistance in Staphylococcus aureus. Indian J Med Res 2008; 127:78-84.

Yoshimasa Y. PCR in Diagnosis of Infection: Detection of Bacteria in Cerebrospinal Fluids. Clin Diagn lab Immunol 2002; 9 (3):508-14.

Markoulatos P, Siafakas N, Moncany M. Multiplex Polymerase Chain Reaction: A Practical Approach. J Clin Lab Anal 2002; 16:47-51.

Elnifro EM, Ashshi AM, Cooper RJ, Klapper PE. Multiplex PCR. Optimization and Application in Diagnostic Virology. Clin Microbiol Rev 2000; 13(4):559-70.

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Published

2017-12-01

How to Cite

Raziq, A. H., Sulaiman, A. M., Merza, N. S., & Abdullah, Z. H. (2017). Molecular detection of methicillin resistant Staphylococcus aureus harbouring β- lactamase resistance genes isolated from different sources of infections. Zanco Journal of Medical Sciences (Zanco J Med Sci), 21(3), 1889–1896. https://doi.org/10.15218/zjms.2017.048

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Original Articles