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Tuon F.F.,Hospital Universitario Evangelico Of Curitiba | Scharf C.,Hospital Universitario Evangelico Of Curitiba | Rocha J.L.,Frischmann Aisengart DASA Medicina Diagnostica | Cieslinsk J.,Hospital Universitario Evangelico Of Curitiba | And 2 more authors.
Brazilian Journal of Infectious Diseases | Year: 2015

Background: Enterobacter is a common nosocomial microorganism and its carbapenem's resistance has increased. The management of these cases is unclear. Objective: We evaluated 16 patients with KPC-producing Enterobacter aerogenes infections, detailing the site of infection, therapy, clinical and epidemiological data. Methods: A retrospective and descriptive study. Clinical data were revised and KPC-2 detection was by molecular methods. Risk factors associated with mortality were compared using appropriate tests according to variable type with a significance level of 0.05. Results: The 30-day mortality rate was 37.5% with no association with inadequate treatment. Age ( p= 0.004) and Charlson score of comorbidities ( p= 0.048) were independent risk factors associated with death in the multivariate analysis. The odds ratio for age >43 years was 3.00 (95% CI: 1.02-9.32) and for Charlson score >3 was 2.00 (95% CI: 1.08-3.71). Five strains were pan-resistant based on automated susceptibility tests. All patients were treated with monotherapy. Conclusion: The clinician should be alert to carbapenem-resistant Enterobacteriaceae infection in older patients with comorbidities. The mortality is high and we believe that prompt and adequate therapy must be employed. © 2015 Elsevier Editora Ltda.

Kottwitz L.B.M.,State University Londrina | Scheffer M.C.,Federal University of Santa Catarina | Dalla-Costa L.M.,Federal University of Parana | Farah S.M.S.S.,Laboratorio Central do Estado LACEN PR | And 3 more authors.
Journal of Medical Microbiology | Year: 2011

A total of 41 Salmonella Enteritidis strains, including phago-types (PTs) PT4 and PT9, were characterized by antimicrobial resistance profiles and PFGE. Of these strains, 34 were isolated from patients and foods, and 7 were of poultry origin. All strains were susceptible to ampicillin, chloramphenicol, cefotaxime, ciprofloxacin and trimethoprim/sulfamethoxazole, and 41.5% (n=17) were resistant to nalidixic acid. PFGE analysis using XbaI and SpeI restriction enzymes resulted in X1S1 as the prevalent pattern, which was present in 48.8% (n=20) of epidemic strains and in one strain isolated from discarded hatching eggs. Distinct patterns were found for the other strains isolated from poultry (X3S1, X8S8, X11S12, X11S13, X16S1 and X13S15). The S. Enteritidis PT9 strains associated with outbreaks of salmonellosis were highly similar (≥0.90), suggesting clonality. The PFGE genotypes were related to the PTs, and it was possible to differentiate strains isolated from patients with salmonellosis from other strains of non-epidemic origin. The PFGE results suggested that the S. Enteritidis strains of poultry origin were a possible source of human salmonellosis during the study period. © 2011 SGM.

Duarte C.A.B.,Instituto Carlos Chagas Fundacao Oswaldo Cruz FIOCRUZ | Duarte C.A.B.,Institute Biologia Molecular do Parana IBMP | Foti L.,Instituto Carlos Chagas Fundacao Oswaldo Cruz FIOCRUZ | Foti L.,Institute Biologia Molecular do Parana IBMP | And 9 more authors.
PLoS ONE | Year: 2010

The strategy used to treat HCV infection depends on the genotype involved. An accurate and reliable genotyping method is therefore of paramount importance. We describe here, for the first time, the use of a liquid microarray for HCV genotyping. This liquid microarray is based on the 59UTR - the most highly conserved region of HCV - and the variable region NS5B sequence. The simultaneous genotyping of two regions can be used to confirm findings and should detect inter-genotypic recombination. Plasma samples from 78 patients infected with viruses with genotypes and subtypes determined in the Versant™ HCV Genotype Assay LiPA (version I; Siemens Medical Solutions, Diagnostics Division, Fernwald, Germany) were tested with our new liquid microarray method. This method successfully determined the genotypes of 74 of the 78 samples previously genotyped in the Versant™ HCV Genotype Assay LiPA (74/78, 95%). The concordance between the two methods was 100% for genotype determination (74/74). At the subtype level, all 3a and 2b samples gave identical results with both methods (17/17 and 7/7, respectively). Two 2c samples were correctly identified by microarray, but could only be determined to the genotype level with the Versant™ HCV assay. Genotype "1" subtypes (1a and 1b) were correctly identified by the Versant™ HCV assay and the microarray in 68% and 40% of cases, respectively. No genotype discordance was found for any sample. HCV was successfully genotyped with both methods, and this is of prime importance for treatment planning. Liquid microarray assays may therefore be added to the list of methods suitable for HCV genotyping. It provides comparable results and may readily be adapted for the detection of other viruses frequently co-infecting HCV patients. Liquid array technology is thus a reliable and promising platform for HCV genotyping. © 2010 Duarte et al.

Nakatani S.M.,Laboratorio Central do Estado LACEN PR | Nakatani S.M.,University of Sao Paulo | Santos C.A.,University of Sao Paulo | Riediger I.N.,Laboratorio Central do Estado LACEN PR | And 8 more authors.
PLoS ONE | Year: 2010

Background: Hepatitis C virus (HCV) genotyping is the most significant predictor of the response to antiviral therapy. The aim of this study was to develop and evaluate a novel real-time PCR method for HCV genotyping based on the NS5B region. Methodology/Principal Findings: Two triplex reaction sets were designed, one to detect genotypes 1a, 1b and 3a; and another to detect genotypes 2a, 2b, and 2c. This approach had an overall sensitivity of 97.0%, detecting 295 of the 304 tested samples. All samples genotyped by real-time PCR had the same type that was assigned using LiPA version 1 (Line in Probe Assay). Although LiPA v. 1 was not able to subtype 68 of the 295 samples (23.0%) and rendered different subtype results from those assigned by real-time PCR for 12/295 samples (4.0%), NS5B sequencing and real-time PCR results agreed in all 146 tested cases. Analytical sensitivity of the real-time PCR assay was determined by end-point dilution of the 5000 IU/ ml member of the OptiQuant HCV RNA panel. The lower limit of detection was estimated to be 125 IU/ml for genotype 3a, 250 IU/ml for genotypes 1b and 2b, and 500 IU/ml for genotype 1a. Conclusions/Significance: The total time required for performing this assay was two hours, compared to four hours required for LiPA v. 1 after PCR-amplification. Furthermore, the estimated reaction cost was nine times lower than that of available commercial methods in Brazil. Thus, we have developed an efficient, feasible, and affordable method for HCV genotype identification. © 2010 Nakatani et al.

Arend L.N.,Federal University of Parana | Arend L.N.,Laboratorio Central do Estado LACEN PR | Pilonetto M.,Laboratorio Central do Estado LACEN PR | de Alencar Siebra C.,Laboratorio Central do Estado LACEN PR | Tuon F.F.,Federal University of Parana
Journal of Infection and Chemotherapy | Year: 2015

Carbapenem-resistant Enterobacteriaceae (CRE) is a major international health problem, and its identification in developing countries is based exclusively on phenotypic methods. The aim of this study was to assess the sensitivity and related parameters of the modified Hodge test (MHT). The assessment was performed in a large number of isolates obtained from different hospitals in several cities of a south Brazilian state. Bacterial species were identified using an automated method. The MHT was performed according to the guidelines set by the CLSI. The gene blaKPC was amplified in order to confirmation CRE expression. The sensitivity, specificity, positive, and negative predictive values were calculated. A total of 942 isolates were submitted to the reference laboratory for confirmation; 143 showed a negative MHT (15.18%) result, while 784 were positive (83.23%), and 15 samples displayed an indeterminate MHT (1.59%) result. All samples expressed the KPC-2 enzyme. Sensitivity, specificity, positive, and negative predictive percentiles were 99%, 89%, 98%, and 99% respectively. We conclude that the modified Hodge test is a reliable test for the prediction of KPC-producing bacteria. © 2014 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases.

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