The identification of Candida species isolated from clinical specimens of immunocompromised patients with PCR and determination of antifungal resistance genes with RFLP and sequencing analysis
Journal Title: Dicle Tıp Dergisi - Year 2012, Vol 39, Issue 2
Abstract
Objectives: The aim of this study is to investigate PCR technique and antifungal resistance genes with RFLP and sequencing analysis in Candida species isolated from clinical specimens of immune-compromised patients. Materials and methods: Clinical samples (96 bronchoalveolar lavages, 56 biopsy-abscess, 8 blood specimens, 15 peritoneal fluid specimens, 15 pleural fluid, 5 cerebrospinal fluid and 5 pericard fluid specimens) from 200 immunosuppressed patients were studied by conventional and molecular methods. Antifungal susceptibility testing was performed by the E-test method. Firstly, fungal DNA was isolated from specimens, and then the resultant products are defined with multiplex PCR. Antifungal resistance and resistance genes were established by E-test and RFLP analysis. Results: Thirty of 200 samples (15%) were culture positive [20 Candida albicans (67%), five Candida parapsilosis (17%), five Candida tropicalis (17%)], and 170 of samples were found culture negative (85%). PCR with the universal primers detected fungal DNA in all 30 culture positive samples. One strain was determined as resistant; 2 strains were dose dependent susceptible and 27 strains were sensitive to fluconazole by E-test. The resistance gene (ERG11) was detected by BamHI and SalI enzymes revealed fluconazole resistance in one of C.albicans strains. The identification was successful in Candida dubliniensis (950 bp) and Candida krusei (360 bp) with multiplex PCR. D132E and E216D mutations were detected in sequencing of ERG 11 gene of this isolate and compared with reference gene in GenBank by clustal analysis. Conclusion: The molecular test methods supplies correct therapy rather early in immunosuppressive patients therefore it is important for the survival.
Authors and Affiliations
Serdar Susever, Yıldız Yeğenoğlu
Keywords
Related Articles
- EP ID EP108780
- DOI -
- Views 127
- Downloads 0
A Case of Neonatal Sepsis due to Listeria monocytogenes Serotype 4b: Case report and literature review
Listeria monocytogenes has been often leads to infection in pregnant women, newborns and patients with impaired immune response. Serotypes 1/2a, 1/2b and 4b are responsible on majority of human infections and outbreaks...
Collagen cross-linking therapy in patients with keratoconus
Keratoconus affecting young population is a progressive eye disease. This disease may remain silent until complain of visual impairment is appeared. By using the high-quality imaging techniques disease now can be diag...
Kronik efüzyonlu otit mediası olan çocuklarda nazal alerjinin semptomatoloji ve deri prick testi ile değerlendirilmesi
Amaç: Bu çalışmada, kronik efüzyonlu otitis media (EOM) bulunan çocuklar, nazal alerji yönünden semptomatolojik olarak ve deri prick testi ile değerlendirilmiş; alerjik rinitin, EOM hastalığı ile birlikteliği ve EOM e...
Single center experience in endovascular aortic repair: review of technical and clinical aspects
Objective: The aim of this study is sharing of our endovascular aortic repair experiments in patients with high risk for surgery and contributing to literature with comparing previous report according to mortality compl...
Behçet hastalığında üst ekstremite trombozu
Behçet hastalığı (BH) tekrarlayan oral, genital ülser ve göz tutulumuyla karakterize, ayrıca eklem, nörolojik, gastrointestinal sistem ve damar tutulumunun da oluşabildiği sistemik bir hastalıktır. BH’ de venöz...