Rapid Detection Of Methicillin Resistant Staphylococci Using Multiplex PCR With Boiling Method For DNA Isolation

  • Ida Bagus Gede Adiguna Wibawa Udayana University
  • Agus Eka Darwinata Udayana University
  • Ni Nengah Dwi Fatmawati Udayana University
  • Nyoman Sri Budayanti Udayana University

Abstract

Methicillin-Resistant Staphylococcus aureus  (MRSA) is Staphylococcus aureus  that has become insusceptible or resistant by


methicillin antibiotic types. Rapid identification of MRSA is essential for early initiation of appropriate antimicrobial therapy.


The aim of this research is to reduce the cost and time needed for multiplex PCR in rapid detection of MRSA by finding an


alternative method for DNA which is boiling method. DNA isolation was performed with boiling method and kit commercial.


The kit method takes time approximately 45 minutes while boiling takes only about 12 minutes. PCR result with boiling


technique used in DNA isolation formed amplification bands of 16S rRNA, mecA , and nuc  in MRSA and 16S rRNA and nuc  in


MSSA. Conclusion can be drawn that boiling method can be used as an alternative method for DNA extraction.

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Author Biographies

Ida Bagus Gede Adiguna Wibawa, Udayana University

Microbiology Departement, Faculty of Medicine 1

Agus Eka Darwinata, Udayana University

Microbiology Departement, Faculty of Medicine 2

Ni Nengah Dwi Fatmawati, Udayana University

Microbiology Departement, Faculty of Medicine 3

Nyoman Sri Budayanti, Udayana University

Microbiology Departement, Faculty of Medicine 4

References

[1] Whitby, M., M. L. McLaws, and G. Berry.2001. Risk of death from
methi- cillin-resistant Staphylococcus aureus bacteraemia: a metaanalysis.
Med. J. Aust. 175:264–267.
[2] Trakulsomboon S, Danchaivijitr S, Rongrungruang Y, Dhiraputra
C,Susaemgrat W, Ito T, Hiramatsu K. First report of methicillin
resistant Staphylococcus aureus reduced susceptibility to vancomycin
in Thailand. J Clin Microbiol. 2001;39:591- 95.
[3] Jan M. Bell and John D. Turnidge. High Prevalence of Oxacillin-
Resistant Staphylococcus aureus Isolates from Hospitalized Patients
in Asia-Pacific and South Africa: Results from SENTRY
Antimicrobial Surveillance Program, 1998- 1999. Antimicrob Agents
Chemother. 2002;46: 879-881.
[4] Vos MC, Ott A,Verbrugh HA. Successful Search-and-Destroy Policy
for Methicillin-Resistant Staphylococcus aureus in The Netherlands
J. Clin.
[5] Microbiol. 2005;43: 2034–2035
[6] acroix, J., K. Jarvi, S. Batra, D. Heritz, and M. Mittelman. 1996.
PCR- based technique for the detection of bacteria in semen and
urine. J. Micro- biol. Methods 26:61–71.
[7] Murakami, K., W. Minamide, K. Wada, E. Nakamura, H. Teraoka,
and S. Watanabe. 1991. Identification of methicillin-resistant strains
of staphylo- cocci by polymerase chain reaction. J. Clin. Microbiol.
29:2240–2244.
[8] Brakstad, O. G., K. Aasbakk, and J. Maeland. 1992. Detection of
Staphylo- coccus aureus by polymerase chain reaction amplification
of the nuc gene. J. Clin. Microbiol. 30:1654–1660.
[9] L. Louie. "Rapid Detection of Methicillin-Resistant Staphylococci
from Blood Culture Bottles by Using a Multiplex PCR Assay",
Journal of Clinical Microbiology, 08/01/2002
Published
2018-01-25
How to Cite
WIBAWA, Ida Bagus Gede Adiguna et al. Rapid Detection Of Methicillin Resistant Staphylococci Using Multiplex PCR With Boiling Method For DNA Isolation. Journal of Health Sciences and Medicine, [S.l.], v. 1, n. 2, p. 8-10, jan. 2018. ISSN 2622-0555. Available at: <https://ojs.unud.ac.id/index.php/jhsm/article/view/36981>. Date accessed: 05 nov. 2024. doi: https://doi.org/10.24843/JHSM.2017.v01.i02.p03.