RESUMO
Presently, several pneumococcal proteins are being evaluated as potential vaccine candidates. Here, we gather novel insights in the immunogenicity of PLY, PsaA, PspA, PspC, NanA, Hyl, PpmA, SlrA, Eno, IgA1-protease, PdBD, BVH-3, SP1003, SP1633, SP1651, SP0189 and SP0376. We developed a multiplex bead-based immunoassay (xMAP(®) Technology, Luminex Corporation) to simultaneously quantify antibodies against these 17 pneumococcal proteins in serum. The median fluorescence intensity (MFI) values obtained for human pooled serum with the multiplex assay were between 82% and 111% (median 94%) of those obtained with the singleplex assays. For IgG, the coefficient of variation (CV) in serum ranged from 2% to 9%, for IgA, the CV ranged from 3% to 14% and for IgM, the CV ranged from 11% to 15%. Using this immunoassay, we showed that anti-pneumococcal antibody levels exhibited extensive inter-individual variability in young children suffering from invasive pneumococcal disease. All proteins, including the proteins with, as yet, unknown function, were immunogenic. In conclusion, the multiplex Streptococcus pneumoniae immunoassay based on proteins is reproducible. This assay can be used to monitor anti-S. pneumoniae antibody responses in a material- and time-saving manner.
Assuntos
Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/imunologia , Infecções Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Proteínas de Bactérias/genética , Criança , Pré-Escolar , Citometria de Fluxo , Humanos , Imunoensaio/métodos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Meningite Pneumocócica/imunologia , Meningite Pneumocócica/microbiologia , Infecções Pneumocócicas/microbiologia , Polissacarídeos Bacterianos/genética , Polissacarídeos Bacterianos/imunologia , Reprodutibilidade dos TestesRESUMO
This prospective study evaluated Raman spectroscopy for the identification of clinically relevant Candida spp. in peritonitis patients. A Raman database was developed by measuring spectra from 93 reference strains belonging to ten different Candida spp. Clinical samples were obtained from the surgical department and intensive care unit of a tertiary university hospital. In total, 88 peritoneal specimens from 45 patients with primary, secondary or tertiary peritonitis were included. Specimens were cultured initially on a selective Sabouraud medium that contained gentamicin to suppress bacterial growth. For conventional identification, a chromogenic medium was used for presumptive identification, followed by use of the Vitek 2 system for definitive identification (requiring a total time of 48-96 h). Raman measurements were taken on overnight cultures from Sabouraud-gentamicin medium. Thirty-one samples were positive for Candida by culture. Using multivariate statistical analyses, a prediction accuracy of 90% was obtained for Raman spectroscopy, which appears to offer an accurate and rapid (12-24 h) alternative for the identification of Candida spp. in peritonitis patients. The reduced turn-around time is of great clinical importance for the treatment of critically ill patients with invasive candidiasis in intensive care units.
Assuntos
Candida/isolamento & purificação , Candidíase/diagnóstico , Peritonite/diagnóstico , Análise Espectral Raman/métodos , Candidíase/microbiologia , Estudos de Viabilidade , Humanos , Peritonite/microbiologia , Estudos Prospectivos , Sensibilidade e Especificidade , Fatores de TempoRESUMO
In the recent years, vibrational spectroscopies (infrared and Raman spectroscopy) have been developed for all sorts of analyses in microbiology. Important features of these methods are the relative ease with which measurements can be performed. Furthermore, in order to obtain infrared or Raman spectra, there is only a limited amount of sample handling involved without the need for expensive chemicals, labels or dyes. Here, we review the potential application of vibrational spectroscopies for the use in medical microbiology. After describing some of the basics of the techniques, considerations on reproducibility and standardisation are presented. Finally, the use of infrared and Raman spectroscopy for the (rapid) identification of medically relevant microorganisms is discussed. It can be concluded that vibrational spectroscopies show high potential as novel methods in medical microbiology.
Assuntos
Técnicas de Tipagem Bacteriana , Candida/classificação , Bactérias Gram-Negativas/classificação , Bactérias Gram-Positivas/classificação , Técnicas de Tipagem Micológica , Infecções Bacterianas/microbiologia , Candidíase/microbiologia , Humanos , Espectrofotometria Infravermelho/métodos , Análise Espectral Raman/métodosRESUMO
BACKGROUND: Short-chain fatty acids (SCFAs) liberated by fermentation of complex carbohydrates might stimulate water and salt absorption, and provide energy. The aim of the study was to assess the number and proportion of faecal bacteria and the concentration of SCFAs of severely malnourished children with cholera receiving oral rehydration solution (ORS) containing glucose, amylase-resistant starch (ARS) or rice. METHODS: Serial faecal samples were collected from 30 malnourished children with cholera until rehydration and partial nutritional recovery. SCFAs were identified and quantitated by high-performance liquid chromatography. In situ hybridization combined with flow cytometry was used to analyse the microbiota in the faeces. RESULTS: Before treatment the concentration of total SCFA in faecal sample of cholera children was found to be 4.7±0.6 mmol/kg and it increased steadily until 95.0±8.7 mmol/kg at day 28. Among different ORS groups, concentration was significantly higher in the Rice-ORS group at day 1 (P<0.011) and at day 2 (P<0.025). During recovery faecal output was significantly reduced and the number of bacteria also increased faster in the Rice-ORS group than in the glucose-ORS group at day 1 and day 2 (P<0.01), and a modest increase in bacterial number was observed in the glucose-ORS plus ARS group (day 1, P=0.07; day 2, P=0.09). CONCLUSION: Clinical recovery was associated with an increase in bacterial and SCFA concentrations with all three carbohydrates in ORS. However, the increases were significantly higher in children receiving Rice-ORS.
Assuntos
Transtornos da Nutrição Infantil/complicações , Cólera/terapia , Ácidos Graxos Voláteis/análise , Fezes/microbiologia , Hidratação/métodos , Transtornos da Nutrição do Lactente/complicações , Soluções para Reidratação/uso terapêutico , Técnicas de Tipagem Bacteriana , Pré-Escolar , Cólera/complicações , Fezes/química , Feminino , Fermentação , Glucose/metabolismo , Glucose/uso terapêutico , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Lactente , Masculino , Oryza/metabolismo , Soluções para Reidratação/química , Soluções para Reidratação/metabolismo , Índice de Gravidade de Doença , Amido/metabolismo , Amido/uso terapêuticoAssuntos
Antibacterianos/uso terapêutico , Infecções por Campylobacter/tratamento farmacológico , Campylobacter fetus/isolamento & purificação , Endocardite Bacteriana/tratamento farmacológico , Antibacterianos/farmacologia , Campylobacter fetus/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Resultado do TratamentoAssuntos
Doença de Crohn/complicações , Nocardiose/complicações , Nocardia asteroides/patogenicidade , Infecções Oportunistas/complicações , Dermatopatias Bacterianas/complicações , Doença de Crohn/tratamento farmacológico , Humanos , Nocardiose/tratamento farmacológico , Nocardia asteroides/classificação , Nocardia asteroides/isolamento & purificação , Infecções Oportunistas/tratamento farmacológico , Filogenia , Dermatopatias Bacterianas/tratamento farmacológicoRESUMO
In this study we aimed to determine the microorganisms found in perianal fistulas in Crohn's disease and whether treatment with ciprofloxacin affects these microorganisms. Thirteen patients (males/females, 7/6; median age, 34 years; range, 18-61 years) with fistulas were treated with infliximab, 5 mg/kg intravenously, at weeks 6, 8, and 12 and randomized to double-blind treatment with ciprofloxacin, 500 mg bd (n = 6), or placebo (n = 7) for 12 weeks. Samples were taken at baseline and at weeks 6 and 18. In the ciprofloxacin group 10 different genera of microorganisms were identified, while 13 genera could be identified in the placebo group. Gram-negative enteric floras were present in a small minority. The genera found in patients with perianal fistulas were predominantly gram-positive microorganisms. Therefore, antimicrobial treatment should be directed toward these microorganisms.
Assuntos
Anti-Infecciosos/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Ciprofloxacina/uso terapêutico , Doença de Crohn/complicações , Fístula Retal/etiologia , Fístula Retal/microbiologia , Pele/microbiologia , Adulto , Infecções por Corynebacterium/tratamento farmacológico , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: The impact of hospitalization on the prevalence of resistant Escherichia coli in the intestinal flora of patients admitted to the surgical wards of three Dutch university-affiliated hospitals was analysed prospectively. METHODS: Faecal samples were obtained on admission to the hospital, at the time of discharge, and 1 and 6 months after discharge. All samples were examined for resistance to nine antibiotic agents. RESULTS: For the total patient population, no significant differences in the prevalence of resistance were observed at the different sampling intervals, except for a significant decrease in cefazolin resistance between the time of discharge and 6 months after discharge (10% to 3%, P < 0.05). This decrease was mainly observed in patients from the university hospital Maastricht (azM), in which a significant decrease from 17% to 6% was detected (P < 0.05). Moreover, despite dissimilarities in patient characteristics and the marked variations in antibiotic use, no significant differences in the prevalence of antibiotic resistance were observed between the three hospitals, except for the overall higher prevalence of cefazolin-resistant E. coli in azM patients (P < 0.05). CONCLUSION: In this study, hospitalization did not seem to have any substantial effect on the prevalence of antibiotic-resistant E. coli at the different time intervals. However, as our study population consisted of surgical patients with a relatively moderate antibiotic use, and the prevalence of antibiotic resistance was only analysed for faecal E. coli, further investigation should be encouraged, as the understanding of the interaction between different resistance reservoirs is important for directing future intervention studies.