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3.
Clin Microbiol Infect ; 26(3): 271-280, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31751768

RESUMO

BACKGROUND: Timely detection of antimicrobial (cephalosporin/carbapenem) resistance (AMR) determinants is crucial to the clinical management of bloodstream infections caused by Gram-negative bacteria (GNB). OBJECTIVES: To review and meta-analyse the evidence for using commercially available molecular tests for the direct detection of AMR determinants in GNB-positive blood cultures (PBCs). DATA SOURCES: PubMed, Scopus and ISI Web of Knowledge. STUDY ELIGIBILITY CRITERIA: Clinical studies evaluating the performance of two major commercial systems, namely the Verigene® and FilmArray® systems, for rapid testing of GNB-PBCs, in comparison with the phenotypic or genotypic methods performed on GNB-PBC isolates. METHODS: Literature search according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses criteria and, for meta-analysis of sensitivity and specificity of both systems, bivariate random-effects model. RESULTS: Twenty studies were identified (3310 isolates) from 2006 to 2019. Nine studies were conducted in East Asia. In 15 studies using phenotypic comparators (1930 isolates), 1014 (52.5%) isolates were Escherichia coli, and 287 (14.9%) of all the isolates displayed AMR phenotypes. In five studies using genotypic comparators (1380 isolates), 585 (42.4%) were E. coli, and 100 (7.2%) of all the isolates displayed AMR genotypes. Pooled sensitivity and specificity estimates for detection of AMR determinants by the Verigene (i.e. CTX-M, IMP, KPC, NDM, OXA and VIM) and/or FilmArray (i.e. KPC) systems were 85.3% (95% CI 79.9%-89.4%) and 99.1% (95% CI 98.2%-99.5%), respectively, across the 15 studies, and 95.5% (95% CI 89.2%-98.2%) and 99.7% (95% CI 99.1%-99.9%), respectively, across the five studies. CONCLUSIONS: Our findings show that the Verigene and FilmArray systems may be a valid adjunct to the conventional microbiology (phenotypic or genotypic) methods used to identify AMR in GNBs. The FilmArray system detects only one AMR genotype, namely KPC, limiting its use. Both Verigene and FilmArray systems can miss important cephalosporin/carbapenem resistance phenotypes in a minority of cases. However, the sensitivity and specificity of both systems render them valuable clinical tools in timely identification of resistant isolates. Further studies will establish the prominence of such rapid diagnostics as standard of care in individuals with bloodstream infections.


Assuntos
Bacteriemia , Farmacorresistência Bacteriana , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/microbiologia , Técnicas de Diagnóstico Molecular , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Anti-Infecciosos , Tomada de Decisão Clínica , Gerenciamento Clínico , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Humanos , Testes de Sensibilidade Microbiana , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
5.
Clin Microbiol Infect ; 24(7): 771-777, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29133157

RESUMO

OBJECTIVES: Biofilm formation (BF) by fungal isolates may dramatically complicate infection. We determined the ability of Candida parapsilosis isolates from single fungaemia episodes to form biofilms and we analysed biofilm subgroups for antifungal susceptibility and pathogenic potential. We then correlated BF with clinical characteristics and outcomes of the episodes. METHODS: BF was measured using the crystal violet biomass assay. Antifungal susceptibility of preformed biofilms was assessed, and virulence was studied using the Galleria mellonella model. A retrospective analysis of patients' clinical records was performed. RESULTS: Of 190 patient-unique isolates, 84, 38 and 68 were identified as having high BF (HBF), moderate BF (MBF) or low BF (LBF), respectively. Among 30 randomly selected isolates, nine (eight HBF and one MBF), six (all HBF) and one (HBF) isolates had elevated sessile minimum inhibitory concentrations to fluconazole, anidulafungin or amphotericin B; all HBF and MBF isolates had elevated voriconazole sessile minimum inhibitory concentrations. G. mellonella killing rates of HBF isolates were significantly greater than MBF (or LBF) isolates (50% vs. 20%, 2 days from infection). By comparing HBF/MBF (106 patients) and LBF (84 patients) groups, we found that HBF/MBF patients had more central venous catheter-related fungaemias (62/106 (58.5%) vs. 29/84 (34.5%), p 0.001) and were more likely to die at 30 days from fungaemia onset (61/106 (57.5%) vs. 28/84 (33.3%), p 0.01). In the HBF/MBF group, azole antifungal therapy and central venous catheter removal were significantly associated with a higher and lower 30-day mortality rate, respectively. CONCLUSIONS: C. parapsilosis BF influences the clinical outcome in patients with fungaemia.


Assuntos
Biofilmes/crescimento & desenvolvimento , Candida parapsilosis/fisiologia , Candida parapsilosis/patogenicidade , Candidemia/microbiologia , Candidemia/mortalidade , Idoso , Idoso de 80 Anos ou mais , Animais , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Biofilmes/efeitos dos fármacos , Bioensaio , Candida parapsilosis/efeitos dos fármacos , Candida parapsilosis/isolamento & purificação , Candidemia/tratamento farmacológico , Infecções Relacionadas a Cateter/tratamento farmacológico , Infecções Relacionadas a Cateter/microbiologia , Infecções Relacionadas a Cateter/mortalidade , Causas de Morte , Feminino , Humanos , Itália , Lepidópteros/microbiologia , Masculino , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Análise de Sobrevida , Virulência
6.
Clin Microbiol Infect ; 23(12): 931-934, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28377311

RESUMO

BACKGROUND: Invasive fungal diseases, including those caused by (multi)drug-resistant Candida and Aspergillus species, still represent global public health concerns. Information about the antifungal susceptibility testing (AFST) of fungal isolates must be quickly produced to help clinicians in administrating appropriate antifungal therapies. Unfortunately, reference AFST methods, albeit accurate, are labour-intensive and take several hours before patients' results can be available to the treating clinicians. AIMS AND SOURCES: This review is a blend of evidence obtained from PubMed literature searches, clinical laboratory experience and the author's opinions that is aimed to summarize recent significant advances and ongoing challenges in the AFST area. CONTENT: Particular attention is given to the new approaches based on genetic or phenotypic recognition of antifungal resistance that are destined to enhance the clinical usefulness of AFST in the near future. Following short-term exposures of fungal cells to antifungal drugs, new antifungal susceptibility end-points have been established, and novel diagnostic assay platforms have been proposed for the genotyping assessment of fungal isolates with resistance-associated mutations. Overall, new approaches provide a rapid, reliable means of identifying those fungal isolates with phenotypically detectable acquired resistance mechanisms, independently from the clinical susceptibility categorization of the isolates as obtained in a classical AFST way. IMPLICATIONS: Despite holding promise as a surrogate diagnostic method to better direct antifungal therapy, the AFST approaches described in this review need to be evaluated in multicentre laboratory studies to enable their standardization and refinement.


Assuntos
Antifúngicos/farmacologia , Testes de Sensibilidade Microbiana , Farmacorresistência Fúngica/genética , Equinocandinas/farmacologia , Genes Fúngicos/genética , Humanos , Testes de Sensibilidade Microbiana/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
7.
Clin Microbiol Infect ; 23(5): 337.e1-337.e3, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28057560

RESUMO

OBJECTIVES: Faecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridium difficile infection (CDI). Although a single faecal infusion is usually sufficient to eradicate CDI, a considerable number of patients need multiple infusions to be cured. The aim of this study was to identify predictors of failure after single faecal infusion in patients with recurrent CDI. METHODS: We included patients with recurrent CDI prospectively treated with FMT by colonoscopy. By means of univariate and multivariate analysis, variables including female gender, age, number of CDI recurrences, severity of CDI, hospitalization, inadequate bowel preparation, unrelated donor, and use of frozen faeces, were assessed to predict failure after single faecal infusion. RESULTS: Sixty-four patients (39 women; mean age 74 years) were included. Of them, 44 (69%) were cured by a single faecal infusion, whereas 20 (31%) needed repeat infusions. Overall, FMT cured 62 of 64 (97%) patients. In the subgroup of patients with severe CDI, only eight of 26 (30%) were cured with a single infusion. At multivariate analysis, severe CDI (OR 24.66; 95% CI 4.44-242.08; p 0.001) and inadequate bowel preparation (OR 11.53; 95% CI 1.71-115.51; p 0.019) were found to be independent predictors of failure after single faecal infusion. CONCLUSIONS: Severe CDI and inadequate bowel preparation appear to be independent predictors of failure after single faecal infusion in patients treated with FMT by colonoscopy for recurrent CDI. Our results may help to optimize protocols and outcomes of FMT in patients with recurrent CDI.


Assuntos
Infecções por Clostridium/terapia , Transplante de Microbiota Fecal , Adulto , Idoso , Idoso de 80 Anos ou mais , Clostridioides difficile , Estudos de Coortes , Colonoscopia , Fezes/microbiologia , Feminino , Microbioma Gastrointestinal , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Recidiva , Falha de Tratamento
8.
Biotechnol Prog ; 32(6): 1584-1591, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27535356

RESUMO

It is now established that the gastric pathogen Helicobacter pylori has the ability to form biofilms in vitro as well as on the human gastric mucosa. The aim of this study is to evaluate the antimicrobial effects of Clarithromycin on H. pylori biofilm and to enhance the effects of this antibiotic by combining it with Alginate Lyase, an enzyme degrading the polysaccharides present in the extracellular polymeric matrix forming the biofilm. We evaluated the Clarithromycin minimum inhibition concentration (MIC) on in vitro preformed biofilm of a H. pylori. Then the synergic effect of Clarithromycin and Alginate Lyase treatment has been quantified by using the Fractional Inhibitory Concentration index, measured by checkerboard microdilution assay. To clarify the mechanisms behind the effectiveness of this antibiofilm therapeutic combination, we used Atomic Force Microscopy to analyze modifications of bacterial morphology, percentage of bacillary or coccoid shaped bacteria cells and to quantify biofilm properties. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1584-1591, 2016.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Claritromicina/farmacologia , Helicobacter pylori/efeitos dos fármacos , Polissacarídeo-Liases/metabolismo , Antibacterianos/química , Claritromicina/química , Relação Dose-Resposta a Droga , Helicobacter pylori/metabolismo , Testes de Sensibilidade Microbiana , Relação Estrutura-Atividade
9.
Eur J Clin Microbiol Infect Dis ; 35(2): 187-93, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26634352

RESUMO

The incidence of Candida bloodstream infections (BSIs) has increased over time, especially in medical wards. The objective of this study was to evaluate the impact of different antifungal treatment strategies on 30-day mortality in patients with Candida BSI not admitted to intensive care units (ICUs) at disease onset. This prospective, monocentric, cohort study was conducted at an 1100-bed university hospital in Rome, Italy, where an infectious disease consultation team was implemented. All cases of Candida BSIs observed in adult patients from November 2012 to April 2014 were included. Patients were grouped according to the initial antifungal strategy: fluconazole, echinocandin, or liposomal amphotericin B. Cox regression analysis was used to identify risk factors significantly associated with 15-day and 30-day mortality. During the study period, 130 patients with candidemia were observed (58 % with C. albicans, 7 % with C. glabrata, and 23 % with C. parapsilosis). The first antifungal drug was fluconazole for 40 % of patients, echinocandin for 57.0 %, and liposomal amphotericin B for 4 %. During follow-up, 33 % of patients died. The cumulative mortality 30 days after the candidemia episode was 30.8 % and was similar among groups. In the Cox regression analysis, clinical presentation was the only independent factor associated with 15-day mortality, and Acute Physiology and Chronic Health Evaluation (APACHE) II score and clinical presentation were the independent factors associated with 30-day mortality. No differences in 15-day and 30-day mortality were observed between patients with and without C. albicans candidemia. In patients with candidemia admitted to medical or surgical wards, clinical severity but not the initial antifungal strategy were significantly correlated with mortality.


Assuntos
Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Candidemia/tratamento farmacológico , Candidemia/mortalidade , Equinocandinas/uso terapêutico , Fluconazol/uso terapêutico , Proteínas Fúngicas/uso terapêutico , Adulto , Idoso , Candida albicans/isolamento & purificação , Candida glabrata/isolamento & purificação , Candidemia/microbiologia , Estudos de Coortes , Feminino , Hospitalização , Humanos , Unidades de Terapia Intensiva , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos Prospectivos , Índice de Gravidade de Doença , Síndrome de Resposta Inflamatória Sistêmica/tratamento farmacológico , Síndrome de Resposta Inflamatória Sistêmica/microbiologia , Síndrome de Resposta Inflamatória Sistêmica/mortalidade
10.
J Clin Microbiol ; 54(3): 576-84, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26677254

RESUMO

Despite the current reliance on blood cultures (BCs), the diagnosis of bloodstream infections (BSIs) can be sped up using new technologies performed directly on positive BC bottles. Two methods (the MALDI BioTyper system and FilmArray blood culture identification [BCID] panel) are potentially applicable. In this study, we performed a large-scale clinical evaluation (1,585 microorganisms from 1,394 BSI episodes) on the combined use of the MALDI BioTyper and FilmArray BCID panel compared to a reference (culture-based) method. As a result, the causative organisms of 97.7% (1,362/1,394) of the BSIs were correctly identified by our MALDI BioTyper and FilmArray BCID-based algorithm. Specifically, 65 (5.3%) out of 1,223 monomicrobial BCs that provided incorrect or invalid identifications with the MALDI BioTyper were accurately detected by the FilmArray BCID panel; additionally, 153 (89.5%) out of 171 polymicrobial BCs achieved complete identification with the FilmArray BCID panel. Conversely, full use of the MALDI BioTyper would have resulted in the identification of only 1 causative organism in 97/171 (56.7%) of the polymicrobial cultures. By applying our diagnostic algorithm, the median time to identification was shortened (19.5 h versus 41.7 h with the reference method; P < 0.001), and the minimized use of the FilmArray BCID panel led to a significant cost savings. Twenty-six out of 31 microorganisms that could not be identified were species/genera not designed to be detected with the FilmArray BCID panel, indicating that subculture was not dispensable for a few of our BSI episodes. In summary, the fast and effective testing of BC bottles is realistically adoptable in the clinical microbiology laboratory workflow, although the usefulness of this testing for the management of BSIs remains to be established.


Assuntos
Sangue/microbiologia , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase/métodos , Sepse/diagnóstico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Algoritmos , Humanos , Técnicas Microbiológicas/economia , Técnicas de Diagnóstico Molecular/economia , Estudos Prospectivos , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/economia , Fatores de Tempo
11.
Clin Microbiol Infect ; 21(5): 493.e1-4, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25698658

RESUMO

The alterations occurring in the intestinal flora during Clostridium difficile infection (CDI) may promote the translocation of Candida to the blood and the development of candidaemia. The aim of our study was to analyse clinical findings of these patients to determine the risk factors associated with the development of candidaemia subsequent to CDI. We compared 35 patients with candidaemia subsequent to CDI with 105 patients with CDI. Patients with candidaemia showed more severe infections and higher mortality. The ribotype 027 strain and vancomycin treatment at ≥ 1000 mg/day were prevalent in patients developing candidaemia. CDI may predispose to the translocation of Candida.


Assuntos
Candidemia/epidemiologia , Clostridioides difficile/isolamento & purificação , Infecções por Clostridium/complicações , Enterocolite/complicações , Idoso , Idoso de 80 Anos ou mais , Antibacterianos/administração & dosagem , Candidemia/tratamento farmacológico , Candidemia/mortalidade , Estudos de Casos e Controles , Clostridioides difficile/classificação , Clostridioides difficile/genética , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/microbiologia , Enterocolite/tratamento farmacológico , Enterocolite/microbiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Ribotipagem , Fatores de Risco , Análise de Sobrevida , Resultado do Tratamento , Vancomicina/administração & dosagem
12.
Int J Immunopathol Pharmacol ; 27(3): 403-12, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25280031

RESUMO

The increased incidence of invasive candidiasis and of patients at risk requires early diagnosis and treatment to improve prognosis and survival. The aim of this study was to set up a ten-protein array-based immunoassay to assess the IgG antibody responses against ten well-known immunogenic C. albicans proteins (Bgl2, Eno1, Pgk1, Pdc11, Fba1, Adh1, Als3, Hwp1, Hsp90 and Grp2) in 51 patients with invasive candidiasis (IC) and in 38 culture-negative controls (non-IC). Antibody levels were higher against Bgl2, Eno1, Pgk1, Als3, Hwp1 and Grp2, than against Adh1, Pdc11, Fba1 and Hsp90, irrespectively of the patient group considered. Moreover, the IgG levels against Bgl2, Eno1, Pgk1 and Grp2 were significantly higher in IC than in non-IC patients. Furthermore, the ROC curves generated by the analysis of the antibody responses against Bgl2, Grp2 and Pgk1 displayed AUC values above 0.7, thus discriminating IC and non-IC patients. According to these results, the employment of the microarray immunoassay (a rapid, sensitive and multiparametric system), in parallel with conventional diagnostics, can help to spot IC patients. This ultimately will allow to initiate an early, focused and optimized antifungal therapy.


Assuntos
Anticorpos Antifúngicos/sangue , Candidíase Invasiva/diagnóstico , Análise Serial de Proteínas/métodos , Proteínas Fúngicas/imunologia , Humanos , Imunoensaio , Imunoglobulina G/sangue , Proteínas Recombinantes/imunologia
13.
Eur J Surg Oncol ; 40(3): 345-51, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24268760

RESUMO

BACKGROUND: Survival of patients after curative surgical resection for gastric cancer (GC) remains poor, thus emphasizing the need for better definition of prognostic factors to improve the long-term course of disease. METHODS: From 1999 to 2009, 110 patients had curative-intent gastrectomy for adenocarcinoma. Clinicopathological features, Helicobacter pylori infection, dietary habits and lifestyle, and the presence of proinflammatory gene polymorphisms were evaluated. RESULTS: At the end of follow-up, 55 deaths had occurred, 48 of them due to GC, whereas the median overall survival (OS) and disease-free survival (DFS) were 62 and 51 months, respectively. From the Kaplan-Meier analysis and log-rank test, statistically significant differences in OS and DFS were found for tumor site (only for DFS), tumor size, lymph node metastasis ratio (NR), and tumor-node-metastasis stage, but not for age, comorbidity, H. pylori infection, cigarette smoking, and IL1B or TNFA polymorphisms. Multivariable Cox regression analysis revealed NR was an independent prognostic factor for OS and DFS. Cardia tumor and patient age 65 years or older were also independent prognostic factors for OS and DFS. CONCLUSIONS: Tumor-related factors remain strongest predictors of survival in GC patients after surgery. Particularly, NR was an effective feature in identifying patients at high risk for adverse outcome.


Assuntos
Adenocarcinoma/mortalidade , Adenocarcinoma/cirurgia , Linfonodos/patologia , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/cirurgia , Adenocarcinoma/secundário , Adulto , Idoso , Estudos de Coortes , Intervalo Livre de Doença , Feminino , Humanos , Itália , Estimativa de Kaplan-Meier , Linfonodos/cirurgia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Invasividade Neoplásica/patologia , Estadiamento de Neoplasias , Prognóstico , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Medição de Risco , Neoplasias Gástricas/patologia , Análise de Sobrevida , Resultado do Tratamento
14.
Microsc Microanal ; 18(5): 1088-94, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23026204

RESUMO

Aspergillus fumigatus has become a leading cause of fungal morbidity and mortality, especially in immunocompromised patients. This fungus is able to grow as a multicellular community and produce a hydrophobic extracellular matrix (ECM), mainly composed of galactomannan and α-1,3 glucans, to protect itself from host defenses and antimicrobial drugs. This matrix envelops the fungus hyphae, binding them into a contiguous sheath on the colony surface, forming a biofilm and increasing the fungal resistance to adverse environmental factors. Adherence to host cells and resistance to physical removal play a key role in fungal colonization and invasion of the host and in a wide range of infections. Here we show that, by using atomic force spectroscopy, it is possible to exploit the peculiar hydrophobicity of the biofilm components (i.e., cell walls, ECM) to detect the biofilm spread, its growth, and lysis on rough surfaces. By means of this approach, we demonstrate that alginate lyase, an enzyme known to reduce negatively charged alginate levels in microbial biofilms, reduces the biofilm adhesion forces suggesting a loss of ECM from the biofilm, which could be used to enhance pharmacological treatments.


Assuntos
Aspergillus fumigatus/enzimologia , Aspergillus fumigatus/crescimento & desenvolvimento , Biofilmes , Proteínas Fúngicas/química , Polissacarídeo-Liases/química , Aspergillus fumigatus/química , Aspergillus fumigatus/citologia , Biofilmes/crescimento & desenvolvimento , Proteínas Fúngicas/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Microscopia de Força Atômica , Polissacarídeo-Liases/metabolismo
15.
Aliment Pharmacol Ther ; 36(3): 222-30, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22650647

RESUMO

BACKGROUND: Helicobacter pylori is one of the most common causes of bacterial infection in humans. Resistance of this infection to conventional therapies has suggested the role of a biofilm-growing bacterium, which is recalcitrant to many antimicrobial agents. AIM: To review the current knowledge on biofilm formation by H. pylori and to discuss the implications of this behaviour in the context of human infections and their treatment. RESULTS: Scanning electron microscopy analysis of gastric biopsies of infected patients demonstrated that H. pylori forms biofilm on the gastric mucosa epithelium. Adaptation to the biofilm environment may produce many persister cells, namely dormant cells, which are highly tolerant to antimicrobials that could account for the recalcitrance of H. pylori infections in vivo. Resistant H. pylori infection has become increasingly common with triple or quadruple therapy, even in the presence of H. pylori strains susceptible to all antibiotics. The mucolytic and thiol-containing antioxidant N-acetylcysteine, associated with antibiotics, was successfully used in clinic for therapy of patients with chronic respiratory tract infections. Consistently, N-acetylcysteine treatment prior to starting antibiotic therapy allowed the disappearance of gastric biofilm in all patients in whom H. pylori was eradicated. CONCLUSION: Effective strategies targeting H. pylori biofilm infections are possible, through the use of substances degrading components of the biofilm.


Assuntos
Antibacterianos/uso terapêutico , Biofilmes/crescimento & desenvolvimento , Mucosa Gástrica/microbiologia , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori/fisiologia , Biofilmes/efeitos dos fármacos , Farmacorresistência Bacteriana , Infecções por Helicobacter/microbiologia , Humanos
16.
Clin Microbiol Infect ; 18(5): 475-84, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-21883662

RESUMO

Accurate species discrimination of filamentous fungi is essential, because some species have specific antifungal susceptibility patterns, and misidentification may result in inappropriate therapy. We evaluated matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification through direct surface analysis of the fungal culture. By use of culture collection strains representing 55 species of Aspergillus, Fusarium and Mucorales, a reference database was established for MALDI-TOF MS-based species identification according to the manufacturer's recommendations for microflex measurements and MALDI BioTyper 2.0 software. The profiles of young and mature colonies were analysed for each of the reference strains, and species-specific spectral fingerprints were obtained. To evaluate the database, 103 blind-coded fungal isolates collected in the routine clinical microbiology laboratory were tested. As a reference method for species designation, multilocus sequencing was used. Eighty-five isolates were unequivocally identified to the species level (≥99% sequence similarity); 18 isolates producing ambiguous results at this threshold were initially rated as identified to the genus level only. Further molecular analysis definitively assigned these isolates to the species Aspergillus oryzae (17 isolates) and Aspergillus flavus (one isolate), concordant with the MALDI-TOF MS results. Excluding nine isolates that belong to the fungal species not included in our reference database, 91 (96.8%) of 94 isolates were identified by MALDI-TOF MS to the species level, in agreement with the results of the reference method; three isolates were identified to the genus level. In conclusion, MALDI-TOF MS is suitable for the routine identification of filamentous fungi in a medical microbiology laboratory.


Assuntos
Aspergillus/classificação , Fusarium/classificação , Mucorales/classificação , Micoses/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Aspergillus/química , Aspergillus/isolamento & purificação , Bases de Dados Factuais , Fusarium/química , Fusarium/isolamento & purificação , Humanos , Mucorales/química , Mucorales/isolamento & purificação , Tipagem de Sequências Multilocus , Técnicas de Tipagem Micológica , Padrões de Referência , Software , Especificidade da Espécie
17.
Clin Microbiol Infect ; 17(1): 44-9, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20132252

RESUMO

As a result of variable expression of biochemical characters, misidentification by conventional phenotypic means often occurs with clinical isolates belonging to Staphylococcus species. Therefore, we evaluated the use of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of 450 blood isolates of the most relevant staphylococcal species, using sequence analysis of the rpoB gene as the reference method. A correct species identification by MALDI-TOF was obtained in 99.3% (447/450), with only three isolates being misidentified. In addition, MALDI-TOF correctly identified all the staphylococcal subspecies studied, including Staphylococcus capitis subsp. capitis and subsp. urealyticus, Staphylococcus cohnii subsp. urealyticus, Staphylococcus hominis subsp. novobiosepticus and subsp. hominis, Staphylococcus saprophyticus subsp. saprophyticus, Staphylococcus schleiferi subsp. schleiferi and Staphylococcus sciuri subsp. sciuri. Thus, MALDI-TOF MS-based species identification of staphylococci can be routinely achieved without any substantial costs for consumables or the time needed for labour-intensive DNA sequence analysis.


Assuntos
Genes Bacterianos/genética , Tipagem Molecular/métodos , Análise de Sequência de DNA/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Infecções Estafilocócicas/microbiologia , Staphylococcus/classificação , Staphylococcus/genética , Especificidade da Espécie
19.
Clin Microbiol Infect ; 16(3): 298-301, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19549221

RESUMO

This study was prospectively conducted in 11 haematology divisions over a 2-year period to evaluate the efficacy of caspofungin in 24 neutropenic patients with haematological malignancies (HM) and candidaemia. These patients had received chemotherapy for HM and were neutropenic (PNN < 0.5 x 10(9)/L) for a median of 12 days (2-41) before candidaemia. The patients received caspofungin for a median duration of 12 days (range 6-26), obtaining a favourable overall response of 58%. At 30 days, 11 patients had died (46%); candidaemia was responsible for mortality in six patients (25%). These results suggest that treatment of candidaemia with caspofungin in neutropenic HM was efficacious, as it is in non-haematological subgroups.


Assuntos
Antifúngicos/uso terapêutico , Candidíase/tratamento farmacológico , Equinocandinas/uso terapêutico , Fungemia/tratamento farmacológico , Neoplasias Hematológicas/complicações , Neutropenia/complicações , Adulto , Idoso , Antineoplásicos/efeitos adversos , Antineoplásicos/uso terapêutico , Candidíase/mortalidade , Caspofungina , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Feminino , Fungemia/mortalidade , Neoplasias Hematológicas/tratamento farmacológico , Humanos , Lipopeptídeos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Resultado do Tratamento , Adulto Jovem
20.
Mycoses ; 53(1): 89-92, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19207832

RESUMO

We report two cases of invasive zygomycoses occurring in severely immunocompromised patients with haematological malignancies that were successfully treated with liposomal amphotericin B and surgical debridement, followed by oral administration of posaconazole. These cases demonstrated that an early instituted, aggressive and combined therapeutic approach results in a recovery from invasive fungal infection, without any relapse of infection, thanks to secondary prophylaxis using posaconazole.


Assuntos
Neoplasias Hematológicas/complicações , Zigomicose/tratamento farmacológico , Zigomicose/cirurgia , Idoso , Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Humanos , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade , Resultado do Tratamento , Triazóis/uso terapêutico
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