Molecular Identification of Burkholderia Cepacia Complex and Species Distribution Among Cystic Fibrosis Patients Seen at the Reference Center in Southern Brazil

Background: Burkholderia cepacia complex (Bcc) infections in cystic fibrosis (CF) patients are associated with decline in lung function and reduced survival. The potential transmissibility of Bcc among CF patients has been reported, indicating that strict segregation of CF patients with Bcc is crucial.Aims: To standardize the PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) assay in order to identify Bcc species and to establish the prevalence of Bcc species and their susceptibility profile among CF patients seen at the Hospital de Clínicas de Porto Alegre (HCPA).Methods: The classification of the clinical isolates recovered from respiratory tract specimens of CF patients as Bcc was achieved using the API-20NE® phenotypic commercial system. The identification of the Bcc species was performed using PCR-RFLP. The antimicrobial disk diffusion susceptibility testing was performed according to the CLSI (2006).Results: API-20NE® was able to identify Bcc isolates (244 specimens), such as B. cepacia, indicating that it was not able to distinguish among the Bcc species. The PCR-RFLP molecular method discriminated the eight reference Bcc species, thus validating the method for clinical isolates. Bcc prevalence determined by PCR-RFLP was 10.6% (26/244). The molecular analysis identified B. cenocepacia in 53.8% (14/26) of infected patients, B. multivorans in 15.4% (4/26), and B. vietnamiensis and B. ambifaria in 7.7% (2/26). The antibiotic resistance profile was variable among Bcc species.Conclusions: The PCR-RFLP method was validated for the identification of Bcc species. B. cenocepacia proved to be the most prevalent species among the CF patients seen at the HCPA

Transforming Knowledge Orders: Museums, Collections and Exhibitions

The history of museums is closely connected not only with the history of collecting and collections, but also with the history of science and humanities. Collections and exhibitions reflect scientific theory and scholarly practice, and in turn shape them. Hence, museums transmit and disseminate, yet also produce knowledge. On the one hand, they visualise and stabilise orders of knowledge through assembling, classifying and fixing objects in exhibitions; on the other hand, new academic paradigms and political changes lead to rearrangements of facts and artefacts in museum storerooms and displays. This volume brings together case studies from various historical and cultural contexts that illuminate such dynamics. Its point of departure is transcultural collections and exhibitions such as cabinets of curiosities and ethnographic collections, whose attempts to inventorise and display the world testify to the desire for, but also the difficulties in establishing and maintaining orders of knowledge. A particular focus is on transformative moments in the history of museums, in particular on the early 1900s, when science and technology museums were established, and on more recent times, which have seen the refurbishment of numerous art and ethnographic museums

Vancomycin resistant Enterococcus spp (VRE) : follow up during 9 years in a tertiary teaching hospital in southern Brazil

Introduction: Infection with vancomycin-resistant Enterococcus spp (VRE) has been a worldwide problem since mid 1980’s and, in Brazil, since 1996. This study was conducted to evaluate the experience with VRE in our institution. Methods: A prospective cohort study from 2000 to 2009 was conducted at Hospital São Lucas da PUCRS. All hospitalized patients with VRE positive culture were included and followed from their diagnosis until they were negative for VRE or their discharge. Only the first admission for each VRE positive patient was included. Pulsed field gel electrophoresis (PFGE) was performed to determine how VRE had spread. Results: A total of 315 cases of VRE were identified, 224 of which were isolated from rectal swabs. Vancomycin-resistant/ampicilin susceptible Enterococcus faecalis were identified in 312 isolates. PFGE was performed in 47 VRE isolates that presented an indistinguishable migratory profile. The median length of hospital stay and length of stay before VRE isolation were 46 days and 21 days, respectively; 52% of the patients were aged 60 and above. The annual distribution of the new VRE cases showed a clear decrease from 2000 to 2009. Discussion: This study shows a substantial VRE colonization (71%) with a homogenous pattern that emphasizes its transversal spread. Predominance of E. faecalis differs from the literature which largely describes a higher prevalence of vancomycin-resistant Enterococcus faecium. The follow up of VRE during 9 years in our institution highlighted the importance of continuous surveillance to prevent outbreaks in our hospital

Tuberculous meningitis: evaluation of polymerase chain reaction (PCR) as a diagnostic tool – a pilot study.

Meningite é uma forma grave e potencialmente fatal de tuberculose. O diagnóstico envolve a detecção de bacilos álcool-ácido resistentes no líquido cefalorraquidiano por microscopia ou cultura. Entretanto, a dificuldade de detectar o organismo representa um desafio ao diagnóstico. O uso da reação em cadeia da polimerase (PCR) na abordagem diagnóstica de meningite causada por Mycobacterium tuberculosis (MTB) tem sido relatado como um método rápido e preciso, com diversos kits comerciais disponíveis. Como alternativa, algumas instituições vêm desenvolvendo testes in house com baixo custo. Em nossa instituição, usamos PCR in house para tuberculose. O desempenho de nossa PCR para o diagnóstico de meningite causada por MTB foi analisado em 148 pacientes consecutivos, usando a cultura do MBT como padrão-ouro. A sensibilidade da PCR no líquido cefalorraquidiano para o diagnóstico de meningite causada por MTB foi de 50%, especificidade de 98,6% e concordância coma cultura de 96% (kappa = 0,52). O desempenho de nossa PCR é semelhante ao obtido com os kits comerciais disponíveis.Meningitis is a severe and potentially fatal form of tuberculosis. The diagnostic workup involves detection of acid-fast bacilli in the cerebrospinal fluid by microscopy or culture. However the difficulty in detecting the organism poses a challenge to diagnosis. Use of polymerase chain reaction (PCR) in the diagnostic approach to Mycobacterium tuberculosis (MTB) meningitis has been reported as a fast and accurate method, with several commercial kits available. As an alternative, some institutions have been developing inexpensive in-house assays. In our institution, we use an in-house PCR for tuberculosis. The performance of our PCR for the diagnosis of MTB meningitis was analyzed in 148 consecutive patients, using MTB culture as the gold standard. Sensitivity of cerebrospinal fluid PCR for the diagnosis of MTB meningitis was 50%, specificity was 98.6%, and concordance with culture was 96% (kappa = 0.52). The performance of our PCR is similar to that obtained with the available commercial kits

Vancomycin resistant enterococcus spp (VRE): follow up during 9 years in a tertiary teaching hospital in southern Brazil

Introduction: Infection with vancomycin-resistant Enterococcus spp (VRE) has been a worldwide problem since mid 1980's and, in Brazil, since 1996. This study was conducted to evaluate the experience with VRE in our institution.Methods: A prospective cohort study from 2000 to 2009 was conducted at Hospital São Lucas da PUCRS. All hospitalized patients with VRE positive culture were included and followed from their diagnosis until they were negative for VRE or their discharge. Only the first admission for each VRE positive patient was included. Pulsed field gel electrophoresis (PFGE) was performed to determine how VRE had spread.Results: A total of 315 cases of VRE were identified, 224 of which were isolated from rectal swabs. Vancomycin-resistant/ampicilin susceptible Enterococcus faecalis were identified in 312 isolates. PFGE was performed in 47 VRE isolates that presented an indistinguishable migratory profile. The median length of hospital stay and length of stay before VRE isolation were 46 days and 21 days, respectively; 52% of the patients were aged 60 and above. The annual distribution of the new VRE cases showed a clear decrease from 2000 to 2009.Discussion: This study shows a substantial VRE colonization (71%) with a homogenous pattern that emphasizes its transversal spread. Predominance of E. faecalis differs from the literature which largely describes a higher prevalence of vancomycin-resistant Enterococcus faecium . The follow up of VRE during 9 years in our institution highlighted the importance of continuous surveillance to prevent outbreaks in our hospital.

Evaluation of identification and susceptibility for Candida spp. isolated directly from positive blood culture bottles

Determination of the susceptibility profile of isolates of Candida from blood culture bottles is extremely important for correctly guiding patient pharmacotherapy. ,e aim of this study was to compare the results of analysis of Candida isolated directly from blood culture bottles by the VITEK MS MALDI-TOF identification system and the fluconazole disk diffusion assay with those of standard identification methods. Testing directly from the bottle allowed results 24 to 48 hours quicker than the standard method. ,ere was a categorical agreement of 51.64% (47 of 91 samples) between the results of analysis directly from the bottle and analysis by the standard method. Regarding species identification, there was 96.15% agreement for Candida parapsilosis (25 of 26 samples). Categorical agreement between the rapid and standard disk diffusion methods was 95%, and the agreement between the rapid disk diffusion method and the broth microdilution method was 97%. Only minor errors in the rapid method were observed: 3 (5%) in the standard disk diffusion method and 2 (3%) in the broth microdilution method. Our study concluded that the rapid disk diffusion method for fluconazole is a fast, easy, reproducible, and consistent method. Its timely implementation for testing antifungal agents in the clinical microbiology laboratory can help reduce profile release times, thus helping to determine the most appropriate antifungal treatment

Meningite tuberculosa : avaliação da reação em cadeia da polimerase como ferramenta diagnóstica : um estudo piloto

Meningitis is a severe and potentially fatal form of tuberculosis. The diagnostic workup involves detection of acidfast bacilli in the cerebrospinal fluid by microscopy or culture. However the difficulty in detecting the organism poses a challenge to diagnosis. Use of polymerase chain reaction (PCR) in the diagnostic approach to Mycobacterium tuberculosis (MTB) meningitis has been reported as a fast and accurate method, with several commercial kits available. As an alternative, some institutions have been developing inexpensive in-house assays. In our institution, we use an inhouse PCR for tuberculosis. The performance of our PCR for the diagnosis of MTB meningitis was analyzed in 148 consecutive patients, using MTB culture as the gold standard. Sensitivity of cerebrospinal fluid PCR for the diagnosis of MTB meningitis was 50%, specificity was 98.6%, and concordance with culture was 96% (kappa = 0.52). The performance of our PCR is similar to that obtained with the available commercial kits.Meningite é uma forma grave e potencialmente fatal de tuberculose. O diagnóstico envolve a detecção de bacilos álcool-ácido resistentes no líquido cefalorraquidiano por microscopia ou cultura. Entretanto, a dificuldade de detectar o organismo representa um desafio ao diagnóstico. O uso da reação em cadeia da polimerase (PCR) na abordagem diagnóstica de meningite causada por Mycobacterium tuberculosis (MTB) tem sido relatado como um método rápido e preciso, com diversos kits comerciais disponíveis. Como alternativa, algumas instituições vêm desenvolvendo testes in house com baixo custo. Em nossa instituição, usamos PCR in house para tuberculose. O desempenho de nossa PCR para o diagnóstico de meningite causada por MTB foi analisado em 148 pacientes consecutivos, usando a cultura do MBT como padrão-ouro. A sensibilidade da PCR no líquido cefalorraquidiano para o diagnóstico de meningite causada por MTB foi de 50%, especificidade de 98,6% e concordância coma cultura de 96% (kappa = 0,52). O desempenho de nossa PCR é semelhante ao obtido com os kits comerciais disponíveis