Molecular epidemiology ofCandidaspecies isolated from clinical specimens of intensive care unit patients

Abstract

Summary Epidemiological analysis of nosocomial Candida infections has gained importance due to an increase in these infections during the recent years. This study investigated the prevalence of clinical infections of Candida in anesthesiology intensive care unit patients, and ascertains the level of genetic diversity in the Candida species. A total of 70 Candida isolates, consisting of 42 Candida albicans, 16 Candida glabrata and 12 Candida tropicalis strains isolated from various clinical sites of infection of anesthesiology intensive care unit patients, were analysed. The susceptibility of the isolates against amphotericin B and fluconazole was determined by microdilution method according to Clinical and Laboratory Standards Institute M27‐A2 standards. The strains were typed by random amplified polymorphic DNA (RAPD)‐PCR using OPE‐03, OPE‐18, RP4‐2 and AP50‐1 primers. In the patients with Candida infections, most isolates exhibited different RAPD patterns. Only three C. albicans pairs isolated within a short time period had the same RAPD pattern. Most of the Candida infections in the anesthesiology intensive care unit of our hospital seem to be caused by endogenous strains. Exogenous spread of C. albicans infections occurred less frequently.

Molecular epidemiology of Candida albicans and Candida glabrata strains isolated from intensive care unit patients in Poland

“…These findings were in accordance with results obtained by da Costa et al (2012) who studied the genetic homology of 15 C. albicans strains using five primers. Similar results were found by Marol and Yücesoy (2008) who used four primers for typing 42 C. albicans strains recovered from ICU patients. Furthermore, four C. albicans isolates cultured from various specimens from the same patients were determined to be different (Marol & Yücesoy 2008).…”

“…Similar results were found by Marol and Yücesoy (2008) who used four primers for typing 42 C. albicans strains recovered from ICU patients. Furthermore, four C. albicans isolates cultured from various specimens from the same patients were determined to be different (Marol & Yücesoy 2008). In our study, the strains CA11 and CA12, isolated from the respiratory tract and urinary system of a single patient, were also unrelated.…”

“…Knowing the source of infection and detection of possible routes of transmission can help with the prevention of clonal spread of pathogenic fungi. RAPD is one of the most frequently used genotyping methods for epidemiological investigations of Candida infections worldwide (Lian et al 2004, Marol & Yücesoy 2008, Muthig et al 2010, Karaman et al 2013). This technique is easy to perform as well as rapid, cost-effective and suitable for the typing of a large number of strains (Noumi et al 2009).…”

“…RAPD reaction - Five arbitrary primers were used for RAPD analysis of C. albicans and C. glabrata strains: CD16AS (5’-CTCTTGAAACTGGGGAGACTTGA-3’), HP1247 (5’-AAGAGCCCGT-3’), ERIC-2 (5’-AAGTAAGTGACTGGGGTGAGCG-3’) (Trojanowska et al 2010), OPE-3 (5’-CCAGATGCAC-3’) and OPE-18 (5’-GGACTGCAGA-3’) (Marol & Yücesoy 2008) (Blirt, Poland). The RAPD reactions were performed with 25 ng of Candida genomic DNA in a final volume of 25 μL containing 100 pmol of primer (Blirt, Poland), 100 μM of each nucleotide (Fermentas, Lithuania), 0.75 mM MgCl 2 , 0.625 U of GoTaq DNA Polymerase in 5 μL and 5x colourless GoTaq Flexi Buffer (pH 8.5) provided by the manufacturer (Promega, USA).…”

Prevalence of Candida Infection at the Intensive Care Unit with Nested Polymerase Chain Reaction (PCR) Using Primer Mixes Specific to Candida DNA Topoisomerase II Genes

“…This indicates reduced susceptibility to fluconazole, as an antifungal agent. Recent studies of nosocomial Candida infections, according to the epidemiologic importance of this phenomenon, have increased (5).…”