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Öğe Evaluation of the Carbapenem Inactivation Method for Detection of Carbapenemase Producing Gram Negative Bacteria in Comparison with the RAPIDEC CARBA NP(Microbial Drug Resistance, 2016) Aktaş, Elif; Malkoçoğlu, Gülşah; Otlu, Barış; Çopur Çiçek, Ayşegül; Külah, Canan; Cömert, Füsun; Sandallı, Cemal; Gürsoy, Nafia Canan; Erdemir, Duygu; Bulut, Mehmet EminTimely detection of carbapenemases by both phenotypic and genotypic methods is essential for developing strategies to control the spread of infections by carbapenem-resistant isolates and related morbidity and mortality. The aim of this study was to compare the performance of a commercial kit, the RAPIDEC® CARBA NP, and an in-house technique, the carbapenem inactivation method (CIM), against a panel of 136 carbapenemase- and noncarbapenemase-producing Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa isolates. RAPIDEC CARBA NP displayed 99% sensitivity and 100% specificity, whereas the sensitivity and specificity were 78% and 100% for the CIM test, respectively. A slight modification of the CIM test, a prolonged incubation time of 4 hours instead of two, increased the sensitivity of the test to 90% by diminishing false negativity particularly for A. baumannii. In conclusion, both tests possess a high performance and are practical for the detection of carbapenemases. Although RAPIDEC CARBA NP is a more rapid and reliable method, the CIM test may represent a useful tool for microbiology laboratories due to its simplicity and availability at any laboratory with low cost.Öğe Femoral Hemodialysis Catheter-Related Bacteremia Due to Globicatella sanguinis: Challenges in Species Identification(Ankara Microbiology Soc, 2017) Aktas, Elif; Gursoy, Nafia Canan; Sakaci, Tamer; Koc, Yener; Hamidi, Aziz Ahmad; Bulut, Emin; Erdemir, DuyguIn this case, catheter-related bacteremia due to Globicatella sanguinis in a 43 years old female patient undergoing hemodialysis with the diagnosis of diabetic nephropathy was presented and the methods in the laboratory diagnosis of the rare opportunistic pathogen, Globicatella cins, were nvestigated. A set of peripheral blood cultures and simultaneous catheter culture was obtained from the patient in third of May 2016. Biochemical tests, Phoenix (Becton Dickinson, USA) and MicroScan (Beckman Coulter, USA) automated systems and matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS) based Microflex MS (Bruker, Daltonics, Germany) and VITEK MS (database v2.0) (bioMerieux, France) systems were used for the identification of the cultured bacteria. Partial 16S rDNA sequencing was done by using specific p8FPL 5'-AGT TTG ATC ATG GCT CAG-3' and p806R 5'-GGA CTA CCA GGG TAT CTA AT-3' primers. Minimal inhibitory concentrations (MICs) for vancomycin, erythromycin, imipenem, cefotaxime and benzypenicillin were determined by agar gradient method. The blood and catheter cultures yielded the same type of bacterial colonies. Alfa-hemolytic, catalase negative colonies observed on blood agar plates after an over night incubation yielded gram-positive cocci on Gram staining. In Sisli Hamidiye Etfal Hospital, the isolate was identifed as G. sulfidifaciens (score value > 2) by Bruker MS system and as G. sanguinis by Phoenix automated system. In Inonu University, the isolate could not be identified by Microscan automated system while VITEK MS named the isolate as 99.9% G. sanguinis and 98.3% G. sulfidifaciens. The 16S rDNA sequencing identifed the isolate as 100% G. sanguinis (GenBank accessionno. KJ680157.1). The MIC values were 0.38 mu g/ml, 1.5 mu g/ml, 0.38 mu g/ ml, > 32 mu g/ml and 64 mu g/ml for vancomycin, eryrthromycin, imipenem, cefotaxime and benzylpenicillin, respectively. The patient was diagnosed as catheter-related bacteremia and vancomycin (1 x 1 g IV/72 h) was used for up to 10 days. No fever and bacterial growth in cultures were present in her control visits. As G. sanguinis is not among the commonly encountered pathogens and due to difficulties in laboratory diagnosis, it may be missedor mis-identified in clinical laboratories. BD Phoenix and Bruker MS data bases lack G. sulfidifaciens and G. sanguinis, respectively, while the Globicatella genus is not present in MicroScan database. The increased number of medical implementations and the increasing number of immunosuppressed patient populations in recenty ears will lead to the emergence of rare bacteria. Increasing the diagnostic power of clinical microbiology laboratories by conventional and molecular methods and renewal of the databases of commercial identification systems by expanding the pathogen spectrum are significantly important for the prevention and control of infections caused by these agents.Öğe Femoral Hemodiyaliz Kateteri ile İlişkili Globicatella sanguinis Bakteremisi: Tür Düzeyinde Tanımlamada Karşılaşılan Sorunlar(2017) Aktaş, Elif; Gürsoy, Nafia Canan; Koç, Yener; Hamidi, Aziz Ahmad; Bulut, Emin; Erdemir, Duygu; Otlu, BarışÖz: Bu olguda diyabetik nefropati tanısıyla kronik hemodiyaliz programına alınan 43 yaşında kadın hastada saptanan Globicatella sanguinis'e bağlı kateterle ilişkili bakteremi olgusu sunulmuştur. Fırsatçı ve nadir bir patojen olan Globicatella cinsinin laboratuvar tanısında kullanılan yöntemler irdelenmiştir. Mayıs 2016 tarihinde hastanın bir set periferik kan kültürü ve eş zamanlı olarak kateter kültürü alınmıştır. Üreyen bakterinin tanımlanması için biyokimyasal testler, Phoenix (BectonDickinson, ABD) ve MicroScan (BeckmanCoulter, ABD) otomatik identifikasyon sistemleri, matriks aracılı lazer desorpsiyon/iyonizasyonuçuş zamanlı kütle spektrometresi (MALDI-TOF MS) temelli Microflex MS (Bruker, Daltonics, Almanya) ve VITEK MS (database v2.0) (bioMérieux, Fransa) sistemleri kullanılmıştır. Etkene özgül p8FPL 5'-AGT TTG ATC ATG GCT CAG-3' ve p806R 5'-GGA CTA CCA GGG TAT CTA AT-3' primerleriyle kısmi 16S rDNA dizi analizi yapılmıştır. Vankomisin, eritromisin, imipenem, sefotaksim ve benzipenisilin için minimum inhibitör konsantrasyonu (MİK) agar gradient yöntemiyle belirlenmiştir. Hastanın kan ve kateter kültürlerinde aynı koloni üremesi tespit edilmiştir. Kanlı besiyerlerinde bir gecelik inkübasyon sonrası gözlenen alfa-hemolitik, katalaz-negatif koloniler, Gram boyama ile gram-pozitif zincir yapan koklar şeklinde görülmüştür. İzolat, Şişli Hamidiye Etfal Eğitim ve Araştırma Hastanesinde; Bruker MS sistemi ile G.sulfidifaciens (skor değeri > 2), Phoenix otomatik identifikasyon sistemi ile G.sanguinis olarak tanımlanmıştır. İnönü Üniversitesi Tıp Fakültesinde; Microscan otomatize sistemiyle tanımlama yapılamamış, VITEK MS ile izolat %99.9 G.sanguinis ve %98.3 G.sulfidifaciens olarak isimlendirilmiştir. 16S rDNA dizi analiziyle izolat %100 Globicatella sanguinis (GenBank accessionno. KJ680157.1) olarak tanımlanmıştır. MİK değerleri vankomisin için 0.38 µg/ml, eritromisin için 1.5 µg/ml, imipenem için 0.38 µg/ml, sefotaksim için > 32 µg/ml ve benzipenisilin için 64 µg/ml olarak belirlenmiştir. Hastada katetere bağlı kan dolaşımı enfeksiyonu olarak düşünüldüğü için, tedaviye vankomisin 1 x 1 g IV/72 saat olarak 10 güne kadar devam edilmiştir. Hastanın kontrollerinde ateş ve üreme olmamıştır. G.sanguinis, sıklıkla karşılaşılan patojenler içerisinde yer almadığından ve laboratuvar tanısında karşılaşılan güçlükler nedeniyle belki de gözden kaçabilmekte veya yanlış tanımlanabilmektedir. BD Phoenix veritabanında G.sulfidifaciens, Bruker MS veritabanında G.sanguinis ve MicroScan veritabanında Globicatella cinsinin mevcut olmadığı görülmüştür. Son yıllarda gelişen tıbbi uygulamalar ve immün sistemi baskılanmış hasta popülasyonundaki artış nedeniyle nadir bakteri türleri daha da sık görülecektir. Klinik mikrobiyoloji laboratuvarlarının, hem klasik mikrobiyolojik yöntemlerle hem de moleküler yöntemlerle tanı gücünün artırılması, ticari identifikasyon sistemi geliştiren şirketlerin patojen spektrumunu genişleterek veritabanlarını yenilemeleri bu tür etkenlerle oluşabilecek ciddi enfeksiyonların önlenmesi açısından önem taşımaktadır.Öğe A First Insight into Escherichia coli ST131 High-Risk Clone Among Extended-Spectrum Beta-Lactamase-Producing Urine Isolates in Istanbul with the Use of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass-Spectrometry and Real-Time PCR(Mary Ann Liebert, Inc, 2017) Aktas, Elif; Otlu, Baris; Erdemir, Duygu; Ekici, Hatice; Bulut, EminObjectives: We aim to investigate, as a first insight, the presence and rates of high-risk Escherichia coli ST131 clone in Istanbul and evaluate antimicrobial resistance and CTX-M-15 production of ST131 and non-ST131 isolates. The use of MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass-spectrometry) to detect E. coli ST131 clone is also evaluated. Materials and Methods: A total of 203 extended-spectrum beta-lactamase (ESBL)-producing urinary isolates from a training hospital in Istanbul were investigated. Detection of E. coli ST131 was done by MALDI-TOF MS and real-time PCR melting curve analysis. The presence of CTX-M and CTX-M-15 beta-lactamases was investigated by PCR and sequence analysis. Results: Of the 203 isolates, 81 (39.9%) and 75 (36.9%) isolates were identified as ST131 clone by PCR and MALDI-TOF MS, respectively. Resistance to ciprofloxacin was significantly higher among ST131 isolates. A total of 169 (83.5%) isolates produced CTX-M beta-lactamase, of which 72 (43%) were CTX-M-15. The production of CTX-M and CTX-M-15 were significantly higher among ST131 isolates. Conclusions: We have demonstrated, for the first time, high rates of ST131 clone among ESBL-producing E. coli isolates in Istanbul, a region with high rates of resistance to third-generation cephalosporins and fluoroquinolones. Further investigation of this high-risk clone and its contribution to high antimicrobial resistance in Turkey is essential. MALDI-TOF MS is a useful tool for detection of high-risk clones and associated resistance patterns, simultaneous to bacterial identification.