Using multiplex PCR as a diagnostic tool to detect methicillin resistant Staphylococcus aureus

Abstract

Aim: The goal of this report was to deliver the methicillin resistant Staphylococcus aureus (MRSA) reports to the clinician and prevent the treatment delays, investigating the efficacy in addition to diagnostic testing and contact isolation strategies for intensive care unit (ICU) patients with MRSA.

Methods: In this report, 320 Staphylococcus aureus strains identified as coagulase positive were cultured from hospitalized ICU patients between 2015 and 2017. Wound swabs were performed and bacteria cultures were evaluated for identification and antibiotic susceptibility testing using a culture antibiogram. Among these cultures from the swabs, MRSA was identified and subsequently screened for the MecA gene using rapid Multiplex polymerase chain reaction (PCR).

Results: MRSA was detected in 67 of 320 strains, because of oxacillin resistance was detected by working with a fully automated culture antibiogram device. In addition, MRSA positivity was detected because of the high MecA gene expression in 56 of these 67 strains using rapid multiplex PCR. 

Conclusion: With greater than 86% sensitivity, patients were able to get early treatment for MRSA due to the rapid screening analysis using Multiplex PCR. This method, as a diagnostic tool, may be of benefit in other diseases.

Keywords

• Staphylococcus aureus,Multiplex PCR,Methicillin resistant Staphylococcus aureus,Wound culture

Metisiline dirençli Staphylococcus aureus'u tespit etmek için bir tanı aracı olarak multipleks PCR kullanılması

Öz

Amaç: Bu çalışmanın amacı, yoğun bakım ünitesi (YBÜ) hastalarında izole edilen, metisiline dirençli Staphylococcus aureus (MRSA) için tanısal test ve temas izolasyon stratejilerine ek olarak etkinliği araştırarak, MRSA raporlarını klinisyene daha hızlı bir şekilde iletmek ve tedavideki gecikmeleri önlemekti.

Yöntemler: Çalışmaya, 2015 ve 2017 yılları arasında, hastaneye yatırılmış YBÜ hastalarından alınan kültür örneklerinden izole edilen ve koagülaz pozitif olarak tanımlanan 320 Staphylococcus aureus suşu dahil edildi. Yara sürüntüleri alındı ve bakteri kültür işlemleri yapılarak, antibiyotik duyarlılık testi ve identifikasyon işlemleri için değerlendirildi. Swablardan alınan bu kültürler arasında MRSA olarak tanımlanan suşlar daha sonra hızlı Multipleks polimeraz zincir reaksiyonu (PCR) kullanılarak MecA geni için tarandı.

Bulgular: 320 suşun 67'sinde, tam otomatik kültür antibiyogram cihazı ile çalışılarak oksasilin direnci saptanarak MRSA tesbit edildi, Ayrıca hızlı Multipleks polimeraz zincir reaksiyonu (PCR) ile çalışıldığında bu 67 suşun 56'sında MecA geni tesbit edilerek, MRSA pozitifliği saptandı.

Sonuç: Multiplex PCR kullanarak hızlı tarama analizine bağlı olarak, % 86'dan fazla duyarlılık ile hastalar MRSA için erken tedavi alabildiler. Bu yöntem, bir tanı aracı olarak, diğer hastalıklarda da yararlı olabilir.

Anahtar Kelimeler

• Staphylococcus aureus,Metisiline dirençli Staphylococcus aureus,Multiplex PCR,Yara kültürü

References

1. 1. Fang H, Hedin G. Rapid Screening and Identification of Methicillin-Resistant Staphylococcus aureus from Clinical Samples by Selective-Broth and Real-Time PCR Assay. J Clin Microbiol. 2003;7;2894–9.
2. 2. Liu Y, Zhang J, and Ji Y. PCR-based Approaches for the Detection of Clinical Methicillin-resistant Staphylococcus aureus. Microbiol J. 2016;14(10):45-56.
3. 3. Deresinski S. Methicillin-resistant Staphylococcus aureus: an evolutionary, epidemiologic, and therapeutic odyssey. Clin Infect Dis. 2005;15;40(4):562-73
4. 4. Carroll KC. Rapid diagnostics for methicillin-resistant Staphylococcus aureus: current status. Mol Diagn Ther. 2008;12(1):15-24.
5. 5. Green BN, Johnson CD, Egan JT, Rosenthal M. Griffith EA, Evans MW. Methicillin-resistant Staphylococcus aureus: an overview for manual therapists. J Chiropr Med. 2012 Mar;11(1):64–76.
6. 6. Köck R, Becker K, Cookson B, van Gemert-Pijnen JE, Harbarth S, Kluytmans J, et al. Methicillin-resistant Staphylococcus aureus (MRSA): burden of disease and control challenges in Europe. Euro Surveill. 2010;41;14:15.
7. 7. Lopez-Alcalde J, Mateos-Mazon M, Guevara M, Conterno LO, Sola I, Cabir Nunes S, et al. Gloves, gowns and masks for reducing the transmission of meticillin-resistant Staphylococcus aureus (MRSA) in the hospital setting. Cochrane Database Syst Rev. 2015;6(7):CD007087.
8. 8. Huletsky A, Giroux R, Rossbach V, Gagnon M, Vaillancourt M, Bernier M, et al. New real-time PCR assay for rapid detection of methicillin-resistant Staphylococcus aureus directly from specimens containing a mixture of staphylococci. J Clin Microbiol. 2004;42(5):1875-84.

9. 9. Bennimath VD, Gavimath CC, Kalburgi PB, Kelmani C. Amplification and Sequencing of MecA Gene From Methicillin Resistance Staphylococcus aureus. Int. J. Adv. Biotechnol. Res. 2011;2(3):310-14.
10. 10. Bakthavatchalam YD, Nabarro LE, Veeraraghavan B. Evolving Rapid Methicillin-resistant Staphylococcus aureus Detection: Cover All the Bases. J Glob Infect Dis. 2017;9(1):18–22.
11. 11. Lucke K, Hombach M, Hug M, Pfyffe GE. Rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) in diverse clinical specimens by the BD GeneOhm MRSA assay and comparison with culture. J Clin Microbiol. 2010;48(3):981-4.
12. 12. Widen R, Healer V, Silbert S. Laboratory Evaluation of the BD MAX MRSA Assay. J Clin Microbiol. 2014;52(7):2686–8.
13. 13. Abbadi S, Yousse H, Nemenqani D, Abdel-Moneim AS. Rapid Identification of Methicillin Resistant Staphylococcus aureus Using Real Time PCR. Advances in Infectious Diseases. 2013;3;44-9.
14. 14. Aqel AA, Alzoubi HM, Vickers A, Pichon B, Kearns AM. Molecular epidemiology of nasal isolates of methicillin-resistant Staphylococcus aureus from Jordan. J Infect Public Health. 2015;8(1):90-7.
15. 15. Hogan B, Rakotozandrindrainy R, Al-Emran H, Dekker D, Hahn A, Jaeger A, Poppert S, et al. Prevalence of nasal colonisation by methicillin-sensitive and methicillin-resistant Staphylococcus aureus among healthcare workers and students in Madagascar. BMC Infect Dis. 2016 Aug 15;16(1):420.
16. 16. Khairalla AS, Wasfi R, Ashour HM. Carriage frequency, phenotypic, and genotypic characteristics of methicillin-resistant Staphylococcus aureus isolated from dental health-care personnel, patients, and environment. Sci Rep. 2017 Aug 7;7(1):7390.