Potential of MALDI-TOF MS as an alternative approach for capsular typing Streptococcus pneumoniae isolates.

Streptococcus pneumoniae can be classified in more than 90 capsular types, as traditionally determined by serological methods and more recently by PCR-based techniques. Such methods, however, can be expensive, laborious or unable to accurately discriminate among certain serotypes. Therefore, determination of capsular types, although extremely important for epidemiological purposes and for estimating the impact of pneumococcal conjugate vaccines, is mainly restricted to research laboratories, being rarely performed in the clinical setting. In the present study, MALDI-TOF MS was evaluated as an alternative tool to characterize 416 pneumococcal isolates belonging to serotypes 6A, 6B, 6C, 9N, 9V or 14. For MALDI-TOF MS analysis, each isolate was submitted to an extraction protocol using formic acid and acetonitrile. Measurements were performed with a Bruker Microflex LT mass spectrometer using default parameters and generating spectra in the range of 2,000-20,000 m/z. Spectra were analyzed with the BioNumerics software v7.6. Isolates were mainly distributed according to the capsular type in a Neighbor Joining tree and serotypes investigated were successfully discriminated by the presence/absence of 14 selected biomarkers. The results suggest that MALDI-TOF MS is a promising alternative for typing pneumococcal strains, highlighting its usefulness for rapid and cost-effective routine application in clinical laboratories.

Práticas de utilização e perfil de contaminação microbiológica de jalecos em escola médica / Use practices and profile of microbiological contaminationof lab coats in medical school

Os jalecos são um veículo potencial de transmissão microbiológica e seu uso inadequado traz riscos à saúde de profissionais e da população. O objetivo deste estudo foi o de conhecer as práticas de utilização e o perfil de contaminação dos jalecos brancos usados como uniforme por docentes e discentes do Centro Universitário Serra dos Orgãos (UNIFESO). Trata-se de um estudo quantitativo, transversal e exploratório. Respeitados os princípios éticos, aplicou-se questionário sobre o modo de uso e se realizou a análise microbiológica. Os dados foram analisados de forma manual, expressos em percentuais e demonstrados por meio de tabelas. Como resultado, 73 (setenta e três) jalecos estudados apresentaram variada flora, com predomínio de cocos Gram positivos. A região do punho, a mais contaminada, apresentou em média 23 UFC por campo. Quanto ao perfil de uso, a maioria, 42, (57%) tem idade entre 18 e 25 anos, 46 (63%) são do sexo feminino, 27 (37%) são acadêmicos de medicina do 3o período e apenas 13 (18%) são docentes. Sobre a frequência de troca do jaleco, 35 (48%) trocam uma vez por semana, entretanto com inadequado manuseio. Após uso, 36 (49%) não lavam o jaleco separadamente, 43 (58%) não usam produto desinfetante e 54 (73%) utilizam a passagem a ferro como medida complementar de contenção microbiológica. Conclui-se que há imperiosa necessidade de conscientização dos acadêmicos e docentes sobre adequados uso e manuseio do jaleco e importância da vestimenta na epidemiologia das infecções intra e extra-ambientes da saúde...

The coats are a potential microbiological transmission means and its misuse brings risks to health professionals and population. The goal of this study was to know the usage practices and the profile of contamination of the smock worn by professors and students of the University Center Serra dos Órgãos (UNIFESO). It is a quantitative study, cross-sectional and exploratory. Respecting ethical principles, a questionnaire on the use of the white coat was applied and the microbiological analysis was held. The data were analyzed manually, expressed in percentages and demonstrated through tables. As a result, 73 (seventy-three) coats studied showed varied flora and predominance of Gram-positive cocci. The region of the wrist, the most contaminated, presented a UFC 23 average per field. As for the usage profile, the majority, 42,(57%) are between the ages of 18 and 25 years, 46 (63%) are female, 32 (44%) are medical students in the 3rd term and only 13 (18%) are professors. On the frequency of changing of lab coat, 35 (48%) change it once a week, however with inappropriate handling. After use, 36 (49%) do not wash the coat separately,43 (58%) do not use disinfectant products, 54 (73%) use ironing as an additional measure against microbiological contamination. It is concluded that there is imperious need for awareness of scholars and professors of appropriate use/handling of the lab coat and the importance of clothing in the epidemiologyof infections intra/extra health care environment...

Emergence of the vanA genotype among Enterococcus gallinarum isolates colonising the intestinal tract of patients in a university hospital in Rio de Janeiro, Brazil.

We describe the characteristics of seven unusual isolates of vancomycin-resistant enterococci (VRE) carrying both the vanC1 and vanA genes that were detected during a 3-month survey carried out to investigate the occurrence of faecal carriage of VRE. The isolates were identified as Enterococcus gallinarum and showed high-level resistance to both vancomycin and teicoplanin (minimum inhibitory concentrations >256 microg/mL and 64-96 microg/mL, respectively). All seven isolates were also resistant to chloramphenicol, erythromycin and high levels of gentamicin, and showed intermediate susceptibility to both quinolones tested (ciprofloxacin and norfloxacin). Susceptibility to fosfomycin, rifampicin and tetracycline varied among isolates. High-level resistance to gentamicin was associated with the aac(6')-aph(2'') gene, and resistance to erythromycin was associated with the erm(B) gene. The seven vanA-carrying E. gallinarum isolates had similar pulsed-field gel electrophoresis (PFGE) profiles. The emergence of multiple antimicrobial resistance, including high-level resistance to glycopeptides, among E. gallinarum points out the need to increase awareness for detection and proper characterisation of these microorganisms, as they may represent potential reservoirs of transmissible, clinically significant resistance genes in nosocomial settings.