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CC BY-NC-ND 4.0 · J Lab Physicians 2012; 4(02): 083-088
DOI: 10.4103/0974-2727.105587
Original Article

Detection of Methicillin Resistance in Staphylococcus Aureus by Polymerase Chain Reaction and Conventional Methods: A Comparative Study

Manju M Pillai
Department of Biochemistry, Subharthi Medical College, Meerut, UP, India
,
Ragunathan Latha
Department of Microbiology, A.V. Medical College, Pondicherry, India
,
Gautam Sarkar
Department of Biochemistry, Subharthi Medical College, Meerut, UP, India
› Institutsangaben Source of Support: Nil.
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ABSTRACT

Background: Methicillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen worldwide, which has emerged over the past 30 years as a leading cause of both nosocomial and community-acquired infections. Accurate and rapid identification of MRSA in clinical specimens is essential for timely decision on effective antimicrobial chemotherapy. Aim: The present study was conducted to compare two conventional phenotypic methods, oxacillin disk diffusion (ODD)

Aim: The present study was conducted to compare two conventional phenotypic methods, oxacillin disk diffusion (ODD) method and mannitol salt agar (MSA) with oxacillin, with polymerase chain reaction (PCR) for mecA gene (as standard).

Materials and Methods: A total of 165 consecutive clinical isolates of S. aureus received at the Department of Microbiology in our tertiary care teaching hospital were included in the study. All the isolates were subjected to ODD (1 μg) method, culture in MSA with oxacillin, and PCR for mecA gene.

Results: The sensitivity and specificity of ODD test were found to be 93.5% (86.4-97.3%) and 83.5% (79.2-85.8%), respectively, and that of MSA with oxacillin were found to be 87.1% (79.5-92.3%) and 89.3% (84.8-92.5%), respectively. The time taken for diagnosing MRSA by conventional methods is 48-72 h, which is more as compared to PCR which takes 18-24 h.

Conclusion: This study recommends advocating PCR for mecA gene on a regular basis for detecting methicillin resistance in S. aureus isolates isolated from sterile body fluids or from special units such as intensive care units.

Keywords

MecA gene - methicillin resistance - polymerase chain reaction - Staphylococcus aureus


Publikationsverlauf

Artikel online veröffentlicht:
09. Mai 2020

© 2012.

Thieme Medical and Scientific Publishers Private Ltd.
A-12, Second Floor, Sector -2, NOIDA -201301, India

 

  • REFERENCES

  • 1 Eveillard M, Martin Y, Hidri N, Boussougant Y, Joly-Guillou ML. Carriage of methicillin-resistant Staphylococcus aureus among hospital employees: Prevalence, duration, and transmission to households. Infect Control Hosp Epidemiol 2004;25:114-20.
  • 2 Ayliffe GA. The progressive international spread of methicillin-resistant Staphylococcus aureus. Clin Infect Dis 1997;24:74-9.
  • 3 Chambers HF. The changing epidemiology of Staphylococcus aureus. Emerg Infect Dis 2001;7:178-82.
  • 4 Timothy R. Peters, Dudley E. Hammon, Rima J. Jarrah, Elizabeth L. Palavecino, Elizabeth S. Blakeney, Katherine A. Poehling. Staphylococcal Toxic Shock Syndrome Complicating Influenza A Infection in a Young Child. ISRN Pulmonology, 2011;2011:1-3.
  • 5 Feudal C, Suvorov M, Vakulenko SB, Mobashery S. The basis for resistance to β-lactam antibiotics by penicillin-binding protein 2a of methicillin-resistant staphylococcus aureus. J Biol Chem 2004;279:40802-6.
  • 6 Shopsin B, Kreiswirth B. Molecular epidemiology of methicillin-resistant Staphylococcus aureus. Emerg Infect Dis 2001;7:322-6.
  • 7 Cosgrove SE, Qi Y, Kaye KS, Harbarth S, Karchmer AW, Carmeli Y et al. The impact of Methicillin Resistance in Staphylococcus aureus Bacteremia on Patient Outcomes: Mortality, Length of Stay, and Hospital Charges. Infect Control Hosp Epidemiol 2005;26:166-74.
  • 8 Cooper BS, Stone SP, Kibbler CC, Cookson BD, Roberts JA, Medley GF, et al. Isolation measures in the hospital management of methicillin resistant Staphylococcus aureus (MRSA): Systematic review of the literature. BMJ 2004;329:533.
  • 9 Towner KJ, Talbot DC, Curran R, Webster CA, Humphreys H. Development and Evaluation of a PCR based immunoassay for the rapid detection of methicillin resistant Staphylococcus aureus. J Med Microbiol 1998;47:607-13.
  • 10 Centers for Disease Control and Prevention (CDC). Staphylococcus aureus resistant to vancomycin. JAMA 2002;288:825.
  • 11 Mathews AA, Thomas M, Appalaraju B, Jayalakshmi J. Evaluation and comparison of tests to detect methicillin resistant S. aureus. Indian J Pathol Microbiol 2010;53:79-82.
  • 12 Brown DF, Edwards DI, Hawkey PM, Morrison D, Ridgway GL, Towner KJ, et al. Guidelines for the laboratory diagnosis and susceptibility testing of methicillin-resistant Staphylococcus aureus (MRSA). J Antimicrob Chemother 2005;56:1000-18.
  • 13 Anand KB, Agrawal P, Kumar S, Kapila K. Comparison of cefoxitin disc diffusion test, oxacillin screen agar, and PCR for mecA gene for detection of MRSA. Indian J Med Microbiol 2009;27:27-9.
  • 14 Van Leeuwen WB, van Pelt C, Luijendijk A, Verbrugh HA, Goessens WH. Rapid Detection of Methicillin Resistance in Staphylococcus aureus Isolates by the MRSA-Screen Latex Agglutination Test. J Clin Microbiol 1999;37:3029-30.
  • 15 CLSI 2011. Performance Standards for antimicrobial susceptibility testing, 21st Informational Supplement. Approved Standard M100-S21. Wayna, pa.
  • 16 Wg Cdr P K Menon, Col A Nagendra. Comparison of rapid method of DNA extraction using microwave irradiation with conventional phenol chloroform technique for use in multiplex PCR for mecA and femB genes. MJAFI 2001;57:194-96.
  • 17 Geha DJ, Uhl JR, Gustaferro CA, Persing DH. Multiplex PCR for the identification of methicillin-resistant Staphylococci in the clinical laboratory. J Clin Microbiol 1994;32:1768-72.
  • 18 Abu Hujier NS, Sharif FA. Detection of methicillin-resistant Staphylococcus aureus in nosocomial infections in Gaza Strip. Afr J Microbiol Res 2008;2:235-41.
  • 19 Swenson JM, Tenover FC. Cefoxitin disk study group. Results of Disk diffusion testing with Cefoxitin correlate with presence of mecA in Staphylococcus sp. J Clin Microbiol. 2005;43:3818-28.
  • 20 Skov R, Smyth R, Clausen M, Larsen AR, Frimodt-Møller N, Olsson-Liljequist B, et al. Evaluation of a cefoxitin30μg disk on Iso-Sensitest agar for detection of methicillin-resistant Staphylococcus aureus. J Antimicrob Chemother 2003;52:204-7.
  • 21 Prasad KN, Kumar R, Tiwari DP, Mishra KK, Ayyagari A. Comparison of various conventional methods with a polymerase chain reaction assay for detecting methicillin resistant and susceptible Staphylococcus aureus strains. Indian J Med Res 2000;112:198-202.
  • 22 Khan AU, Sultan A, Tyagi A, Zahoor S, Akram M, Kaur S. Amplification of mecA gene in multi-drug resistant Staphylococcus aureus strains from hospital personnel. J Infect Dev Ctries 2007;3:289-95.
  • 23 Lewis RA, Dyke KG. MecI represses synthesis from the beta-lactamase operon of Staphylococcus aureus. J Antimicrob Chemother 2000;45:139-44.
  • 24 Fluit AC, Wielders CL, Verhoef J, Schmitz FJ. Epidemiology and susceptibility of 3,051 Staphylococcus aureus isolates from 25 university hospitals participating in the European SENTRY study . J Clin Microbiol 2001;39:3727-32.
  • 25 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.