25S Intron analysis followed by restriction enzyme digestion performed for genotyping Candida albicans isolates

Candida albicans, özellikle bağışıklığı baskılanmış konaklarda enfeksiyon etkeni olarak en sık izole edilen fungal patojendir. Uygun enfeksiyon kontrol stratejilerinin geliştirilmesi ve epidemiyolojik veri toplanması açısından, enfeksiyon etkeni olan mikroorganizmaların genotiplendirilmesi önem taşımaktadır. 25S intron analizi, C.albicans suşlarının genotiplendirilmesinde kullanılan kolay ve güvenilir bir yöntem olarak kabul edilmektedir. Bununla birlikte, ayırım gücünün düşük olması, epidemiyolojik araştırmalarda bu yöntemin kullanımını sınırlandırmaktadır. Bu çalışmada, enfeksiyon etkeni olarak izole edilen C.albicans suşlarının polimeraz zincir reaksiyonu (PCR) ile 25S intron analizini takiben, elde edilen PCR ürünlerinin restriksiyon enzim analizine tabi tutulmasıyla ayırım gücü daha yüksek olan bir genotiplendirme sistemi oluşturulması amaçlanmıştır. Bu çalışmaya, çeşitli klinik örneklerden (121 kan, 69 balgam, 36 vajinal akıntı, 26 yara, 8 idrar) enfeksiyon etkeni olarak izole edilmiş 260 adet C.albicans suşu dahil edilmiştir. Tüm izolatlara, PCR ile literatürde belirtildiği şekilde 25S intron analizi uygulanmış ve elde edilen PCR ürünleri HaeIII restriksiyon enzimi ile kesilerek genotiplendirilmiştir. Her iki yöntemin ayırım gücü ayrı ayrı hesaplanmıştır. 25S intron analizi ile 184 (%70.8) izolat genotip A, 42 (%16.2) izolat genotip B ve 34 (%1 izolat da genotip C olarak bulunmuş; yöntemin ayırım gücü 0.46 olarak hesaplanmıştır. PCR ürünlerinin HaeIII ile kesimi sonucunda genotip A’da 10, genotip B’de bir ve genotip C’de beş ayrı kesim paterni (genotip) tespit edilmiş; böylece restriksiyon enzim analizi eklenmesiyle elde edilen genotip sayısı 16’ya, yöntemin ayırım gücü de 0.79’a yükselmiştir. Farklı genotiplendirme yöntemlerinin birlikte kullanımı, genotip sayısını artırarak ayırım gücünü yükseltmekle birlikte, aynı suşların farklı genotiplere dağılmasıyla sonuçlanabilmekte ve bu durum değerlendirmede güçlüklere neden olabilmektedir. 25S intron analizini takiben HaeIII restriksiyon enzim analizi uygulanması ise, tamamen farklı bir genotiplendirme yöntemi eklenmesine gerek kalmaksızın yöntemin ayırım gücünü yükseltmekte ve yöntemi epidemiyolojik araştırmalar ve klinik izolatların genotiplendirilmesi için daha uygun hale getirmektedir. HaeIII yerine başka enzimlerin kullanılması yöntemin ayırım gücünü daha da yükseltebilir. Bu yöntemle elde edilen genotiplerle hasta özellikleri, klinik veriler, antifungal duyarlılıklar gibi parametreler arasında ilişki olup olmadığını araştırmak için daha detaylı araştırmalar planlanabilir.Candida albicans is the most frequently encountered fungal pathogen especially in the immunocompromised hosts. Genotyping clinical microbial isolates is important for obtaining epidemiological data and for establishing appropriate infection control strategies in the hospital setting. 25S intron analysis is an easy and reliable method used for genotyping C.albicans strains. As it has a low discriminatory power, its use is limited in epidemiological studies. In this study, our aim was to genotype clinical C.albicans isolates by using 25S intron analysis followed by restriction enzyme digestion in order to develop a more discriminative genotyping system for C.albicans . A total of 260 clinical C.albicans strains isolated from various infection sites (121 blood, 69 sputum, 36 vaginal discharge, 26 wound, 8 urine samples) were genotyped by 25S intron analysis, and all the products obtained by polymerase chain reaction (PCR) were digested with HaeIII restriction enzyme. Discriminatory power of each method was calcula- ted. Among the isolates 184 (70.8%) were classified as genotype A, 42 (16.2%) as genotype B, and 34 (13%) as genotype C by 25S intron analysis. Discriminatory power of the method was calculated as 0.46. HaeIII restriction of genotype A, B and C isolates produced ten, one, and five restriction patterns (genotypes), respectively. By the addition of restriction enzyme analysis, the number of genotypes obtained was increased to 16, and the discriminatory power of the method to 0.79. Combining different genotyping methods increases the discriminatory power by increasing the number of genotypes obtained. However, there is also a risk to split certain strains in different genotypes by the different methods used and this makes the genotypic evaluation more difficult. On the other hand, combining 25S intron analysis with restriction enzyme analysis increases the discriminatory power without introducing a totally different method, and makes the method more suitable for epidemiological purposes and for genotyping clinical isolates. Different enzymes instead of HaeIII should be tested to evaluate the effect on the discriminatory power. In order to evaluate the relationship between the genotypes obtained by this method and parameters such as patient characteristics, clinical data, and antifungal susceptibilities, more sophisticated studies can be performed

Resistance to newer beta-Lactams and related ESBL types in gram-negative nosocomial isolates in Turkish hospitals: Results of the multicentre HITIT study

Gram-negatif hastane izolatlarında geniş spektrumlu (B-laktamazlar (GSBL) ve çeşitli direnç mekanizmaları nedeniyle B-laktam antibiyotiklere karşı giderek artan direnç, bu antibiyotiklerin ampirik kullanımını sınırlamaktadır. Tedaviye yön vermek için çok merkezli sürveyans çalışmalarına gereksinim vardır. Türkiye'de altı merkezin katılımı ile Haziran 2004-0cak 2005 tarihleri arasında yapılan çok merkezli bu çalışmada, 457 Escherichia coli, 390 Klebsiella pneumoniae, 194 Pseudomonas aeruginosa ve 155 Acineto-bacter baujvannii olmak üzere toplam 1196 gram-negatif hastane izolatının seftriakson, seftazidim, sefe-pim, imipenem, sefoperazon/sulbaktam ve piperasilin/tazobaktama karşı in vitro direnç durumunun ve ayrıca E.coli ve K.pneumoniae kan izolatlarında GSBL varlığının araştırılması amaçlanmıştır. Antibiyotiklerin minimum inhibisyon konsantrasyonu (MİK) değerleri ve GSBL üretimi E-test (AB Biodisk, Solna) yöntemiyle saptanmıştır. Direnç oranlarının belirlenmesinde CLSI direnç sınır değerleri kullanılmış; GSBL türleri PCR analizi ile belirlenmiştir. Çalışmada, E.coli suşlarının %26'sında, K.pneumoniae suşlarının ise %32'sinde GSBL pozitifliği saptanmış; bu oranlar kan izolatı olan E.coli ve K.pneumoniae suşlarında sırasıyla %31.7 ve %33.3 olarak izlenmiştir. Kan izolatlarında en sık tespit edilen enzimler, CTX-M (%71.4), TEM (%49.4) ve SHV (%46.7) türevleri olmuş; en sık saptanan CTX-M türünün CTX-M-15 (%69.4) olduğu, bunu CTX-M-3 (%28.6) ve CTX-M-1 (%2)'in izlediği görülmüştür. E.coli izolatlarında imipeneme direnç saptanmazken, K.pneumoniae1da imipenem direnci %1.3, P.aeruginosa'da %28.9, A.baumannii'de ise %52.2 olarak bulunmuştur. E.coli, K.pneumoniae, P.aeruginosa ve A.baumannii'de sefoperazon/sulbak-tama karşı direnç oranları sırasıyla %6, %17.7, %27.9 ve %4J .3 olarak belirlenmiş, piperasilin/tazobak-tam direnci ise sırasıyla %10.2, %22.3, %22.7 ve %78.7 olarak bulunmuştur. Bu sonuçlar, ülkemizin çeşitli merkezlerinde hastane izolatı gram-negatif bakterilerde GSBL oranlarının ve R-laktamlara direncin yüksek olduğunu, merkezler arasında direnç yönünden farklılıklar olduğunu ve ülkemizde CTX-M enzimlerinin hastane izolatlarında yaygın olduğunu göstermiştir.Increasing resistance due to extended-spectrum beta-lactamases (ESBLs) and multiple resistance mechanisms in gram-negative hospital isolates restrict the role of beta-lactam antibiotics in empirical treatment of serious infections. As the prevalence of ESBL producing strains and resistance rates to antimicrobial agents can vary in each center, local surveillance studies are required to guide therapy. In this study, in vitro rates of resistance to ceftriaxone, ceftazidime, cefepime, imipenem, cefoperazone/sulbactam and piperacillin/tazobactam were evaluated in 1196 gram-negative hospital isolates in a multicenter in vitro study with the participation of six different centers in Turkey between the period of June 2004-January 2005. The isolates included Escherichia coli (n= 457), Klebsiella pneumoniae (n= 390), Pseudomonas aeruginosa (n= 194) and Acinetobacter boumannii (n= 155). In addition, frequency of ESBL production and types of enzymes were determined in blood isolates of E. coli and K. pneumoniae. MICs and ESBL production were investigated by E-test (AB Biodisk, Solna) and the results were evaluated by using CLSI breakpoints. PCR analysis was used for typing of the ESBLs. In E. coli, 26% and in K. pneumoniae 32% of the isolates were ESBL producers. Among the blood isolates of E. coli and K. pneumoniae, 31.7% and 33.3% produced ESBLs, respectively. CTX-M (71.4%) was the most prevalent enzyme, followed by TEM (49.4%) and SHV (46.7%) derived enzymes. CTX-M-15 (69.4%) was the most frequent CTX-M type in blood isolates followed by CTX-M-3 (28.6%) and CTX-M-1 (2%). Resistance to imipenem was not observed in E. coli isolates, however it was 1.3% in K. pneumoniae, 28.9% in P. aeruginosa and 52.2% in A. baumannii strains. Resistance to cefoperazone/sulbactam was found as 6%, 17.7%, 27.9% and 41.3% in E. coli, K. pneumoniae, P. aeruginosa and A. baumannii isolates, respectively, whereas resistance rates to piperacillin/tazobactam were 10.2%, 22.3%, 22.7% and 78.7%, respectively. These results indicate that ESBL production and rates of resistance to beta-lactam antibiotics are high in hospital isolates of gram-negative bacteria in Turkey, however, they show variations in different hospitals and CTX-M enzymes are prevalent in these isolates

Impact of a multidimensional infection control approach on central line-associated bloodstream infections rates in adult intensive care units of 8 cities of Turkey: findings of the International Nosocomial Infection Control Consortium (INICC)

Geyik, Mehmet Faruk/0000-0002-0906-0902; Leblebicioglu, Hakan/0000-0002-6033-8543; Dikmen, Yalim/0000-0002-3122-5099; Unal, Necmettin/0000-0002-9440-7893WOS: 000320023600001PubMed: 23641950Background: Central line-associated bloodstream infections (CLABs) have long been associated with excess lengths of stay, increased hospital costs and mortality attributable to them. Different studies from developed countries have shown that practice bundles reduce the incidence of CLAB in intensive care units. However, the impact of the bundle strategy has not been systematically analyzed in the adult intensive care unit (ICU) setting in developing countries, such as Turkey. The aim of this study is to analyze the impact of the International Nosocomial Infection Control Consortium (INICC) multidimensional infection control approach to reduce the rates of CLAB in 13 ICUs of 13 INICC member hospitals from 8 cities of Turkey. Methods: We conducted active, prospective surveillance before-after study to determine CLAB rates in a cohort of 4,017 adults hospitalized in ICUs. We applied the definitions of the CDC/NHSN and INICC surveillance methods. The study was divided into baseline and intervention periods. During baseline, active outcome surveillance of CLAB rates was performed. During intervention, the INICC multidimensional approach for CLAB reduction was implemented and included the following measures: 1-bundle of infection control interventions, 2-education, 3-outcome surveillance, 4-process surveillance, 5-feedback of CLAB rates, and 6-performance feedback on infection control practices. CLAB rates obtained in baseline were compared with CLAB rates obtained during intervention. Results: During baseline, 3,129 central line (CL) days were recorded, and during intervention, we recorded 23,463 CL-days. We used random effects Poisson regression to account for clustering of CLAB rates within hospital across time periods. The baseline CLAB rate was 22.7 per 1000 CL days, which was decreased during the intervention period to 12.0 CLABs per 1000 CL days (IRR 0.613; 95% CI 0.43 - 0.87; P 0.007). This amounted to a 39% reduction in the incidence rate of CLAB. Conclusions: The implementation of multidimensional infection control approach was associated with a significant reduction in the CLAB rates in adult ICUs of Turkey, and thus should be widely implemented.Foundation to Fight against Nosocomial InfectionsThe funding for the activities carried out at INICC head quarters were provided by the corresponding author, Victor D. Rosenthal, and Foundation to Fight against Nosocomial Infections

Impact of a multidimensional infection control approach on catheter-associated urinary tract infection rates in adult intensive care units in 10 cities of Turkey: International Nosocomial Infection Control Consortium findings (INICC)

Yalcin, Ata Nevzat/0000-0002-7243-7354; dursun, oguz/0000-0001-5482-3780; Geyik, Mehmet Faruk/0000-0002-0906-0902; Dursun, Oguz/0000-0001-5482-3780; Leblebicioglu, Hakan/0000-0002-6033-8543; Unal, Necmettin/0000-0002-9440-7893WOS: 000325254600011PubMed: 23623158Background: We evaluate the effectiveness of a multidimensional infection control approach for the reduction of catheter-associated urinary tract infections (CAUTIs) in 13 intensive care units (ICUs) in 10 hospital members of the International Nosocomial Infection Control Consortium (INICC) from 10 cities of Turkey. Methods: A before-after prospective active surveillance study was used to determine rates of CAUTI. The study was divided into baseline (phase 1) and intervention (phase 2). In phase 1, surveillance was performed applying the definitions of the Centers for Disease Control and Prevention/National Healthcare Safety Network. In phase 2, we implemented a multidimensional approach that included bundle of infection control interventions, education, surveillance and feedback on CAUTI rates, process surveillance, and performance feedback. We used random effects Poisson regression to account for clustering of CAUTI rates across time periods. Results: The study included 4,231 patients, hospitalized in 13 ICUs, in 10 hospitals, in 10 cities, during 49,644 patient-days. We recorded a total of 41,871 urinary catheter (UC)-days: 5,080 in phase 1 and 36,791 in phase 2. During phase 1, the rate of CAUTI was 10.63 per 1,000 UC-days and was significantly decreased by 47% in phase 2 to 5.65 per 1,000 UC-days (relative risk, 0.53; 95% confidence interval: 0.4-0.7; P value = .0001). Conclusion: Our multidimensional approach was associated with a significant reduction in the rates of CAUTI in Turkey. Copyright (C) 2013 by the Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.Foundation to Fight against Nosocomial InfectionsFunding for the activities carried out at INICC headquarters were provided by the corresponding author, Victor D. Rosenthal, and the Foundation to Fight against Nosocomial Infections

Impact of the International Nosocomial Infection Control Consortium (INICC) Multidimensional Hand Hygiene Approach, over 8 years, in 11 cities of Turkey

Aims: To evaluate the effectiveness of the International Nosocomial Infection Control Consortium (INICC) Multidimensional Hand Hygiene Approach in Turkey and analyse predictors of poor hand hygiene compliance. Design: An observational, prospective, interventional, before-and-after study was conducted from August 2003 to August 2011 in 12 intensive care units (ICU) of 12 hospitals in 11 cities. The study was divided into a baseline and a follow-up period and included random 30-minute observations for hand hygiene compliance in ICU. The hand hygiene approach included administrative support, supplies availability, education and training, reminders in the workplace, process surveillance, and performance feedback. Results: We observed 21,145 opportunities for hand hygiene. Overall hand hygiene compliance increased from 28.8% to 91% (95% CI 87.6–93.0, p 0.0001). Multivariate and univariate analyses showed that several variables were significantly associated with poor hand hygiene compliance: males vs. females (39% vs. 48%; 95% CI 0.79–0.84, p 0.0001), ancillary staff vs. physicians (35% vs. 46%, 95% CI 0.73–0.78, p 0.0001), and adult vs. pediatric ICUs (42% vs. 74%, 95% CI 0.54–0.60, p 0.0001). Conclusions: Adherence to hand hygiene was significantly increased with the INICC Hand Hygiene Approach. Specific programmes should be directed to improve hand hygiene in variables found to be predictors of poor hand hygiene compliance

Impact of the International Nosocomial Infection Control Consortium (INICC) Multidimensional Hand Hygiene Approach, over 8 years, in 11 cities of Turkey

AIMS: To evaluate the effectiveness of the International Nosocomial Infection Control Consortium (INICC) Multidimensional Hand Hygiene Approach in Turkey and analyse predictors of poor hand hygiene compliance. DESIGN: An observational, prospective, interventional, before-and-after study was conducted from August 2003 to August 2011 in 12 intensive care units (ICU) of 12 hospitals in 11 cities. The study was divided into a baseline and a follow-up period and included random 30-minute observations for hand hygiene compliance in ICU. The hand hygiene approach included administrative support, supplies availability, education and training, reminders in the workplace, process surveillance, and performance feedback. RESULTS: We observed 21,145 opportunities for hand hygiene. Overall hand hygiene compliance increased from 28.8% to 91% (95% CI 87.6–93.0, p 0.0001). Multivariate and univariate analyses showed that several variables were significantly associated with poor hand hygiene compliance: males vs. females (39% vs. 48%; 95% CI 0.79–0.84, p 0.0001), ancillary staff vs. physicians (35% vs. 46%, 95% CI 0.73–0.78, p 0.0001), and adult vs. pediatric ICUs (42% vs. 74%, 95% CI 0.54–0.60, p 0.0001). CONCLUSIONS: Adherence to hand hygiene was significantly increased with the INICC Hand Hygiene Approach. Specific programmes should be directed to improve hand hygiene in variables found to be predictors of poor hand hygiene compliance