Murat ARAL, Hacer UĞURLU, Esra KAYA, Kezban Tülay YALÇINKAYA, Zerife ORHAN

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

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.

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

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

PDF

___

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 meticillinresistant 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 methicillinresistant Staphylococcus aureus and glycopeptideresistant 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, MRSASelect 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.