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Çeşitli Klinik Örneklerden Izole Edilen Staphylococcus aureus Suşlarında Metisilin Direncinin Araştırılmasında Konvansiyonel PCR Yöntemi ile Sefoksitin Disk Difüzyon, Otomatize Sistem ve Chromogenic MRSA Agar Yöntemlerinin Karşılaştırılması

Year 2019, Volume: 22 Issue: 3, 451 - 455, 30.06.2019
https://doi.org/10.18016/ksutarimdoga.vi.457495

Abstract

Metisiline dirençli Staphylococcus
aureus
(MRSA) en önemli enfeksiyöz ajanlardan biridir. Bu nedenle MRSA'nın
hızlı ve doğru tanısı çok önemlidir. PCR ile mecA gen tespiti referans
metodu olmakla birlikte, basit olmasından dolayı rutin uygulamada geleneksel
yöntemler tercih edilmektedir. Geleneksel yöntemler ile tanının koyulması 48-96
saat sürdüğü için çeşitli kromojenik besiyerleri geliştirilmiştir.
Çalışmamızdaki amacımız metisilin direncinin tespitinde kullanılan çeşitli yöntemleri
PCR ile karşılaştırmaktır. Çeşitli klinik örneklerden izole edilen 48 S.
aureus
suşu çalışmamıza dahil edilmiştir.48 S.aureus suşunun 19'u mecA
gen-pozitif ve 29'u negatiftir. mecA gen-pozitif 19 suş da otomatik
sistem ve disk difüzyonu ile dirençli bulunmuştur.  Kromojenik agarda, 19
MRSA suşunun 15'i metisilin dirençli ve 4'ü duyarlı bulunmuştur.29 mecA
gen-negatif suş, otomatik sistem ve disk difüzyonu ile duyarlı bulunmuştur. mecA
geni negatif olan 29 MSSA suşunun kromojenik agarda 17’si dirençli olarak
değerlendirmiştir.

References

  • Broekema NM, Van TT, Monson TA, Marshall SA, Warshauer DM 2009. Comparison of cefoxitin and oxacillin disk diffusion methods for detection of mecA-mediated resistance in Staphylococcus aureus in a large-scale study. J Clin Microbiol. 47: 217-219.
  • Cesur S, Altın N, Yapar Toros G, Koldaş K, Solgun G, Şencan İ 2014. Çeşitli Klinik Örneklerden İzole Edilen Staphylococcus aureus Suşlarında Metisilin Direncinin Belirlenmesinde Chrom-ID MRSA Besiyerinin Değerlendirilmesi. Orta Doğu Tıp Dergisi. 6(2): 80-85.
  • Cesur S, Yildiz E, Irmak H, et al. 2010. Evaluation of oxacillin resistance screening agar and chromogenic MRSA agar media for the detection of methicillin resistance in Staphylococcus aureus clinical isolates. Mikrobiyol Bul. 44(2): 279-284.
  • Clinical and Laboratory Standarts Institute 2014. Performance Standards for Antimicrobial Susceptibility Testing. 24th Informational Supplement, M100-S24, Clinical and Laboratory Standards Institute, Wayne, PA 19087 USA.
  • Data P, Gulati N, Singla N et al. 2011. Evaluation of various methods for the detection of meticillin-resistant Staphylococcus aureus strains and susceptibility patterns. J Med Microbiol. 60(Pt 11): 1613-1616.
  • DeLeo FR, Chambers HF 2009. Reemergence of antibiotic-resistant Staphylococcus aureus in the genomics era. J Clin Invest. 119(9): 2464-2474.
  • Denys GA, Renzi PB, Koch KM, Wissel CM 2013. Three-way comparison of BBL CHROMagar MRSA II, MRSA Select, and spectra MRSA for detection of methicillin resistant Staphylococcus aureus isolates in nasal surveillance cultures. J Clin Microbiol. 51(1): 202-205.
  • Feng Y, Chen CJ, Su LH, Hu S, Yu J et al. 2008. Evolution and pathogenesis of Staphylococcus aureus lessons larned from genotyping and comparative genomics. FEMS Microbiol Rev. 32: 23-37.
  • Kluytmans J 2007. Control of methicillin resistant Staphylococcus aureus (MRSA) and the value of rapid tests. J Hosp Infect. 65(S2): 100-104.
  • Lodise TP, McKinnon PS 2005. Clinical and economic impact of methicillin resistance in patients with Staphylococcus aureus bacteremia. Diagn. Microbiol. Infect. Dis. 52: 113-122.
  • Malhotra-Kumar, S, Haccuria K, Michiels M, Ieven M, Poyart C, Hryniewicz W, Goossens H 2008. Current trends in rapid diagnostics for methicillin-resistant Staphylococcus aureus and glycopeptide-resistant enterococcus species. J. Clin. Microbiol. 46: 1577-1587.
  • Marlowe EM, Bankowski MJ 2011. Conventional and molecular methods for the detection of methicillinresistant Staphylococcus aureus. J Clin Microbiol. 49: 53-56.
  • Nahimana I, Francioli P, Blanc DC 2006. Evaluation of three chromogenic media (MRSA-ID, MRSA-Select and CHROMagar MRSA) and ORSAB for surveillance cultures of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect. 12(12): 1168-1174.
  • Özen NS, Dağlar D, Baysan BÖ, Yıldırım Ç, Yazısız H, Öğünç D, Öngüt G, Çolak D, Gültekin M 2011. Metisilin dirençli Staphylococcus aureus suşlarının saptanmasında MRSA ID kromojenik besiyerinin değerlendirilmesi. ANKEM Derg. 25(1): 31-34.
  • Paule SM, Hacek DM, Kufner B et al. 2007. Performance of the BD GeneOhm methicillin resistant Staphylococcus aureus test before and during high-volume clinical use. J Clin Microbiol. 45(9): 2993-2998.
  • Perry JD and Freydiere AM 2007. The application of chromogenic media in clinical microbiology. J. Appl. Microbiol. 103:2046–2055.
  • Prosper M, Veras N, Azarian T, Rathore M, Nolan D et al. 2013. Molecular epidemiology of community assicuated methicillin-resistant Staphylococcus aureus in the genomic era: a cross-sectional study. Sci Rep. 3: 1-8.
  • Roisin S, Nonhoff C, Denis O, Struelens M 2008. Evaluation of new Vitek 2 card and disk diffusion method for determining susceptibility of Staphylococcus aureus to oxacillin. J Clin Microbiol. 46: 2525-2528.
  • Strommenger B, Kettlitz C, Werner G, Witte W. 2003. Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol. 41(9): 4089-4094.
  • Struelens MJ 2006. Rapid identification of methicillin resistant Staphylococcus aureus (MRSA) and patient management. Clin Microbiol Infect. 12 (suppl 9): 23-26.
  • Sürücüoğlu S, Sakarya M, Gazi H, Ecemiş T, Kurutepe S 2011. Riskli hastalarda metisiline dirençli Staphylococcus aureus taşıyıcılığının belirlenmesinde hızlı tanı testlerinin değerlendirilmesi. Turk Hij Den Biyol Derg. 68(3): 115-121.
  • Uzun B, Karataş Şener AG, Güngör S, Afşar İ, Yüksel Ergin Ö, Demirci M 2013. Staphylococcus aureus Suşlarındaki Metisilin Direncinin Belirlenmesinde Sefoksitin Disk Difüzyon Testi, Otomatize Sistem ve Kromojenik Besiyerinin Karşılaştırılması. Mikrobiyol Bul. 47(1): 11-18.
  • Van Hal SJ, Stark D, Lockwood B, Marriott D, Harkness J 2007. Methicillin resistant Staphylococcus aureus (MRSA) detection: comparison of two molecular methods (IDI-MRSA PCR assay and GenoType MRSA Direct PCR assay) with three selective MRSA agars (MRSA ID, MRSASelect, and CHROMagar MRSA) for use with infection-control swabs. J Clin Microbiol. 45(8): 2486-2490.
  • Von Eiff C, Maas D, Sander G, Friedrich AW, Peters G, Becker K 2008. Microbiological evaluation of a new growthbased approach for rapid detection of methicillin-resistant Staphylococcus aureus. J Antimicrob Chemother. 61(6): 1277-1280.
  • Xu Z, Hou Y, Peters BM, Chen D, Li B, Li L, Shirtliff ME 2016. Chromogenic media for MRSA diagnostics. Molecular Biol Rep J. 43(11): 1205-1212.
  • Zhu LX, Zhang ZW, Wang C, Yang HW, Zhang Q, Cheng J 2006. Evaluation of the CLSI cefoxitin 30-microg disk-diffusion method for detecting methicillin resistance in staphylococci. Clin Microbiol Infect. 12(10): 1039-1042.

Comparison of Conventional PCR Method With Cefoxitin Disc Diffusion, Automated System and Isolation on Chromogenic MRSA Medium Methods for The Detection of Meticillin Resistance In Staphylococcus aureus Strains Isolated From Various Clinical Specimens

Year 2019, Volume: 22 Issue: 3, 451 - 455, 30.06.2019
https://doi.org/10.18016/ksutarimdoga.vi.457495

Abstract

Meticillin-resistant Staphylococcus
aureus
(MRSA) is one of the most important infectious agents. Therefore,
fast and accurate diagnosis of MRSA is utmost important. Although mecA
gene detection by PCR is reference method, conventional methods are preferred
in routine practices due to simplicity. As conventional methods last 48-96
hours, several chromogenic media have been developed. Our objective was to
compare the methods used for meticillin resistance detection with PCR.
Forty-eight S. aureus strains isolated from various clinical specimens
were included. Of the 48 S. aureus strains, 19 were mecA gene-positive
and 29 were negative. mecA gene-positive 19 strains were also
meticillin-resistant by automated system and disk diffusion. On chromogenic
agar, 15 of 19 MRSA strains were meticillin-resistant and 4 were
meticillin-sensitive. Twentynine mecA gene-negative strains were
susceptible to meticillin by automatized system and disk diffusion. Among 29 mecA
gene-negative MSSA strains inoculated on chromogenic agar, 17 were
methicillin-resistant. According to our study, chromogenic media would be
ineffective to detected to meticillin-resistance because of low sensitivity and
specificity in routine.

References

  • Broekema NM, Van TT, Monson TA, Marshall SA, Warshauer DM 2009. Comparison of cefoxitin and oxacillin disk diffusion methods for detection of mecA-mediated resistance in Staphylococcus aureus in a large-scale study. J Clin Microbiol. 47: 217-219.
  • Cesur S, Altın N, Yapar Toros G, Koldaş K, Solgun G, Şencan İ 2014. Çeşitli Klinik Örneklerden İzole Edilen Staphylococcus aureus Suşlarında Metisilin Direncinin Belirlenmesinde Chrom-ID MRSA Besiyerinin Değerlendirilmesi. Orta Doğu Tıp Dergisi. 6(2): 80-85.
  • Cesur S, Yildiz E, Irmak H, et al. 2010. Evaluation of oxacillin resistance screening agar and chromogenic MRSA agar media for the detection of methicillin resistance in Staphylococcus aureus clinical isolates. Mikrobiyol Bul. 44(2): 279-284.
  • Clinical and Laboratory Standarts Institute 2014. Performance Standards for Antimicrobial Susceptibility Testing. 24th Informational Supplement, M100-S24, Clinical and Laboratory Standards Institute, Wayne, PA 19087 USA.
  • Data P, Gulati N, Singla N et al. 2011. Evaluation of various methods for the detection of meticillin-resistant Staphylococcus aureus strains and susceptibility patterns. J Med Microbiol. 60(Pt 11): 1613-1616.
  • DeLeo FR, Chambers HF 2009. Reemergence of antibiotic-resistant Staphylococcus aureus in the genomics era. J Clin Invest. 119(9): 2464-2474.
  • Denys GA, Renzi PB, Koch KM, Wissel CM 2013. Three-way comparison of BBL CHROMagar MRSA II, MRSA Select, and spectra MRSA for detection of methicillin resistant Staphylococcus aureus isolates in nasal surveillance cultures. J Clin Microbiol. 51(1): 202-205.
  • Feng Y, Chen CJ, Su LH, Hu S, Yu J et al. 2008. Evolution and pathogenesis of Staphylococcus aureus lessons larned from genotyping and comparative genomics. FEMS Microbiol Rev. 32: 23-37.
  • Kluytmans J 2007. Control of methicillin resistant Staphylococcus aureus (MRSA) and the value of rapid tests. J Hosp Infect. 65(S2): 100-104.
  • Lodise TP, McKinnon PS 2005. Clinical and economic impact of methicillin resistance in patients with Staphylococcus aureus bacteremia. Diagn. Microbiol. Infect. Dis. 52: 113-122.
  • Malhotra-Kumar, S, Haccuria K, Michiels M, Ieven M, Poyart C, Hryniewicz W, Goossens H 2008. Current trends in rapid diagnostics for methicillin-resistant Staphylococcus aureus and glycopeptide-resistant enterococcus species. J. Clin. Microbiol. 46: 1577-1587.
  • Marlowe EM, Bankowski MJ 2011. Conventional and molecular methods for the detection of methicillinresistant Staphylococcus aureus. J Clin Microbiol. 49: 53-56.
  • Nahimana I, Francioli P, Blanc DC 2006. Evaluation of three chromogenic media (MRSA-ID, MRSA-Select and CHROMagar MRSA) and ORSAB for surveillance cultures of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect. 12(12): 1168-1174.
  • Özen NS, Dağlar D, Baysan BÖ, Yıldırım Ç, Yazısız H, Öğünç D, Öngüt G, Çolak D, Gültekin M 2011. Metisilin dirençli Staphylococcus aureus suşlarının saptanmasında MRSA ID kromojenik besiyerinin değerlendirilmesi. ANKEM Derg. 25(1): 31-34.
  • Paule SM, Hacek DM, Kufner B et al. 2007. Performance of the BD GeneOhm methicillin resistant Staphylococcus aureus test before and during high-volume clinical use. J Clin Microbiol. 45(9): 2993-2998.
  • Perry JD and Freydiere AM 2007. The application of chromogenic media in clinical microbiology. J. Appl. Microbiol. 103:2046–2055.
  • Prosper M, Veras N, Azarian T, Rathore M, Nolan D et al. 2013. Molecular epidemiology of community assicuated methicillin-resistant Staphylococcus aureus in the genomic era: a cross-sectional study. Sci Rep. 3: 1-8.
  • Roisin S, Nonhoff C, Denis O, Struelens M 2008. Evaluation of new Vitek 2 card and disk diffusion method for determining susceptibility of Staphylococcus aureus to oxacillin. J Clin Microbiol. 46: 2525-2528.
  • Strommenger B, Kettlitz C, Werner G, Witte W. 2003. Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol. 41(9): 4089-4094.
  • Struelens MJ 2006. Rapid identification of methicillin resistant Staphylococcus aureus (MRSA) and patient management. Clin Microbiol Infect. 12 (suppl 9): 23-26.
  • Sürücüoğlu S, Sakarya M, Gazi H, Ecemiş T, Kurutepe S 2011. Riskli hastalarda metisiline dirençli Staphylococcus aureus taşıyıcılığının belirlenmesinde hızlı tanı testlerinin değerlendirilmesi. Turk Hij Den Biyol Derg. 68(3): 115-121.
  • Uzun B, Karataş Şener AG, Güngör S, Afşar İ, Yüksel Ergin Ö, Demirci M 2013. Staphylococcus aureus Suşlarındaki Metisilin Direncinin Belirlenmesinde Sefoksitin Disk Difüzyon Testi, Otomatize Sistem ve Kromojenik Besiyerinin Karşılaştırılması. Mikrobiyol Bul. 47(1): 11-18.
  • Van Hal SJ, Stark D, Lockwood B, Marriott D, Harkness J 2007. Methicillin resistant Staphylococcus aureus (MRSA) detection: comparison of two molecular methods (IDI-MRSA PCR assay and GenoType MRSA Direct PCR assay) with three selective MRSA agars (MRSA ID, MRSASelect, and CHROMagar MRSA) for use with infection-control swabs. J Clin Microbiol. 45(8): 2486-2490.
  • Von Eiff C, Maas D, Sander G, Friedrich AW, Peters G, Becker K 2008. Microbiological evaluation of a new growthbased approach for rapid detection of methicillin-resistant Staphylococcus aureus. J Antimicrob Chemother. 61(6): 1277-1280.
  • Xu Z, Hou Y, Peters BM, Chen D, Li B, Li L, Shirtliff ME 2016. Chromogenic media for MRSA diagnostics. Molecular Biol Rep J. 43(11): 1205-1212.
  • Zhu LX, Zhang ZW, Wang C, Yang HW, Zhang Q, Cheng J 2006. Evaluation of the CLSI cefoxitin 30-microg disk-diffusion method for detecting methicillin resistance in staphylococci. Clin Microbiol Infect. 12(10): 1039-1042.
There are 26 citations in total.

Details

Primary Language English
Journal Section RESEARCH ARTICLE
Authors

Esra Kaya 0000-0002-0732-6471

Murat Aral 0000-0002-3576-4380

Zerife Orhan 0000-0003-2154-3074

Kezban Tülay Yalçınkaya 0000-0002-6324-4585

Hacer Uğurlu 0000-0001-6126-5502

Publication Date June 30, 2019
Submission Date September 5, 2018
Acceptance Date January 30, 2019
Published in Issue Year 2019Volume: 22 Issue: 3

Cite

APA Kaya, E., Aral, M., Orhan, Z., Yalçınkaya, K. T., et al. (2019). Comparison of Conventional PCR Method With Cefoxitin Disc Diffusion, Automated System and Isolation on Chromogenic MRSA Medium Methods for The Detection of Meticillin Resistance In Staphylococcus aureus Strains Isolated From Various Clinical Specimens. Kahramanmaraş Sütçü İmam Üniversitesi Tarım Ve Doğa Dergisi, 22(3), 451-455. https://doi.org/10.18016/ksutarimdoga.vi.457495


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