Production of extended-spectrum beta-lactamases (ESBL) by enterobacteria is an important resistance mechanism against antimicrobial beta-lactamics. We tested 498 bacterial strains isolated from two tertiary-care teaching hospitals for ESBL production, using screening breakpoints for aztreonam and third generation cephalosporins, according to CLSI recommendations. Among these isolates, 155 were positive for the ESBL screening test, and 121 (78%) were confirmed by the clavulanic acid combination disk method. We found a high frequency of ESBL (24%) among Enterobacteriaceae, with a frequency of 57.4% for Klebsiella pneumoniae, 21.4% for Klebsiella oxytoca, and 7.2% for E. coli. In other members of Enterobacteriaceae, non-Klebsiella and non-E. coli, the prevalence was 21.6%. Ceftriaxone and cefotaxime showed a higher sensitivity in the screening test (99.2%) when compared to ceftazidime, aztreonam and cefpodoxime. However, cefotaxime/cefotaxime plus clavulanic acid showed a higher sensitivity in the confirmatory test (96.7%).

The predisposition of patients with cystic fibrosis (CF) for recurrent pulmonary infections can result in poor prognosis of the disease. Although the clinical significance in CF of microorganisms, such as Staphylococcus aureus, Haemophilus influenzae and Pseudomonas aeruginosa, is well established, the implication of uncommon glucose non-fermenting Gramnegative bacilli (UGNF-GNB) in respiratory samples from CF patients is still unclear. Because of limitations of traditional methods used in most clinical laboratories, the accurate identification of these microbes is a challenge. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) is an alternative tool for efficient identification of bacteria. This was a retrospective study to evaluate different identification methods in a collection of UGNF-GNB isolated from children with CF during a period of three years. The performance of MALDI-TOF was compared to that of 16S rDNA gene sequencing and to a conventional and automated phenotypic identification. The discriminatory power of MALDI-TOF (75.0 % agreement) was superior to automated techniques (67.1 % agreement) and to conventional phenotypical identification (50.0 % agreement). MALDI-TOF also demonstrated high accuracy in identifying Stenotrophomonas maltophilia, Achromobacter xylosoxidans and Chryseobacterium indologenes, but had limited utility in identifying Pandoraea spp. and some species of Acinetobacter and Chryseobacterium (other than C. indologenes). Although MALDI-TOF identified only 75 % of the isolates in comparison with 16S rDNA gene sequencing, the prompt identification and high discriminatory power exhibited by MALDI-TOF make it a useful tool for the characterization of micro-organisms that are difficult to identify using routine methods.

Cystic fibrosis (CF) patients with Burkholderia cepacia complex (Bcc) pulmonary infections have high morbidity and mortality. The aim of this study was to compare different methods for identification of Bcc species isolated from paediatric CF patients. Oropharyngeal swabs from children with CF were used to obtain isolates of Bcc samples to evaluate six different tests for strain identification. Conventional (CPT) and automatised (APT) phenotypic tests, polymerase chain reaction (PCR)-recA, restriction fragment length polymorphism-recA, recAsequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) were applied. Bacterial isolates were also tested for antimicrobial susceptibility. PCR-recA analysis showed that 36 out of the 54 isolates were Bcc. Kappa index data indicated almost perfect agreement between CPT and APT, CPT and PCR-recA, and APT and PCR-recA to identify Bcc, and MALDI-TOF and recAsequencing to identify Bcc species. The recAsequencing data and the MALDI-TOF data agreed in 97.2% of the isolates. Based on recA sequencing, the most common species identified were Burkholderia cenocepacia IIIA (33.4%),Burkholderia vietnamiensis (30.6%), B. cenocepaciaIIIB (27.8%), Burkholderia multivorans (5.5%), and B. cepacia (2.7%). MALDI-TOF proved to be a useful tool for identification of Bcc species obtained from CF patients, although it was not able to identify B. cenocepacia subtypes.

artigo de reVisão review article A Gardnerella vaginalis e as infecções do trato urinário The Gardnerella vaginalis and the urinary tract infections

Introdução: Na maioria dos laboratórios de Microbiologia considera-se contaminação uma cultura de urina com mais de um morfotipo colonial, ignorando-se o desenvolvimento ou solicitando-se novo material. Raramente o isolado é considerado significativo. Objetivos: Com a finalidade de estudar as infecções urinárias polimicrobianas, no período de agosto de 2003 a janeiro de 2004, na cidade de Tubarão, Santa Catarina, foram selecionadas 117 amostras de urina de pacientes internados no Hospital Nossa Senhora da Conceição, de ambos os sexos, com idades que variavam de 14 a 98 anos. Métodos: Realizou-se uma análise minuciosa dos prontuários dos pacientes. Procedeu-se o Gram da gota de urina não centrifugada e a cultura com alça calibrada de 1 ou 10 μl em ágar CLED (cistina-lactose deficiente em eletrólitos). Todas as culturas foram repetidas com nova coleta de urina com supervisão direta. Os clínicos aguardaram a segunda coleta (confirmatória) para iniciar a antibioticoterapia. Descartaram-se os pacientes que iniciaram a antibioticoterapia imediatamente após a primeira coleta ou que estavam utilizando antimicrobianos. Resultados: Obteve-se o total de seis (12,8%) culturas polimicrobianas confirmadas, em um universo de 47 amostras com crescimento bacteriano estudadas. Os resultados foram compatíveis com as indicações clínicas. Conclusão: É importante ressaltar a correlação entre os achados laboratoriais e as indicações clínicas dos pacientes. Recomenda-se avaliar criteriosamente isolados polimicrobianos em amostras de urina, sejam ambulatoriais ou hospitalares. resumo unitermos Infecção urinária polimicrobiana Gram de urina não centrifugada Urocultura polimicrobiana

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