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1.
Front Cell Infect Microbiol ; 11: 634215, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34381737

RESUMO

Bloodstream infections (BSIs), the presence of microorganisms in blood, are potentially serious conditions that can quickly develop into sepsis and life-threatening situations. When assessing proper treatment, rapid diagnosis is the key; besides clinical judgement performed by attending physicians, supporting microbiological tests typically are performed, often requiring microbial isolation and culturing steps, which increases the time required for confirming positive cases of BSI. The additional waiting time forces physicians to prescribe broad-spectrum antibiotics and empirically based treatments, before determining the precise cause of the disease. Thus, alternative and more rapid cultivation-independent methods are needed to improve clinical diagnostics, supporting prompt and accurate treatment and reducing the development of antibiotic resistance. In this study, a culture-independent workflow for pathogen detection and identification in blood samples was developed, using peptide biomarkers and applying bottom-up proteomics analyses, i.e., so-called "proteotyping". To demonstrate the feasibility of detection of blood infectious pathogens, using proteotyping, Escherichia coli and Staphylococcus aureus were included in the study, as the most prominent bacterial causes of bacteremia and sepsis, as well as Candida albicans, one of the most prominent causes of fungemia. Model systems including spiked negative blood samples, as well as positive blood cultures, without further culturing steps, were investigated. Furthermore, an experiment designed to determine the incubation time needed for correct identification of the infectious pathogens in blood cultures was performed. The results for the spiked negative blood samples showed that proteotyping was 100- to 1,000-fold more sensitive, in comparison with the MALDI-TOF MS-based approach. Furthermore, in the analyses of ten positive blood cultures each of E. coli and S. aureus, both the MALDI-TOF MS-based and proteotyping approaches were successful in the identification of E. coli, although only proteotyping could identify S. aureus correctly in all samples. Compared with the MALDI-TOF MS-based approaches, shotgun proteotyping demonstrated higher sensitivity and accuracy, and required significantly shorter incubation time before detection and identification of the correct pathogen could be accomplished.


Assuntos
Bacteriemia , Infecções Estafilocócicas , Bacteriemia/diagnóstico , Candida albicans , Escherichia coli , Humanos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Infecções Estafilocócicas/diagnóstico , Staphylococcus aureus
2.
Med Mycol ; 58(4): 485-492, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31504817

RESUMO

The gut microbiota harbor a wide range of bacterial species, but also yeasts may be part of this ecosystem. Infants who are being treated in intensive care units are often colonized by Candida species. However, little is known regarding commensal yeast colonization of healthy infants and young children. Here the acquisition of yeast species was studied in a birth-cohort including 133 healthy Swedish infants. A rectal swab sample was obtained on day 3 of life, and fresh fecal samples were obtained at regular intervals up to 3 years of age; the samples were cultured quantitatively for yeasts. Colonization with yeasts increased rapidly in the first months of life, with 73/133 infants (55%) colonized at 6 months of age. The yeast numbers in positive samples decreased from an average of 105 cfu/g in infants aged 0-2 months to 103.5 cfu/g at 3 years of age. Candida albicans was the most frequently isolated species and reached higher population counts than the other species in culture-positive infants. The yeast colonization rate did not differ between infants who were delivered vaginally and those birthed via Caesarean section, whereas breastfed infants showed a lower colonization rate (p < 0.05 for 1 year of age compared to the other infants). The results demonstrate that yeasts, particularly C. albicans and C. parapsilosis (sensu lato), are common commensals in the gut microbiota of healthy infants and young children.


Assuntos
Candida/fisiologia , Candidíase/microbiologia , Portador Sadio/microbiologia , Fezes/microbiologia , Microbioma Gastrointestinal , Simbiose , Candida/classificação , Candida/isolamento & purificação , Candida albicans/isolamento & purificação , Cesárea , Pré-Escolar , Parto Obstétrico , Feminino , Voluntários Saudáveis , Humanos , Lactente , Recém-Nascido , Estudos Longitudinais , Masculino , Suécia
3.
Mycoses ; 62(11): 1043-1048, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31376228

RESUMO

The aim of this study was to estimate the annual burden of fungal infections in Sweden using data mainly from 2016. Data on specific populations were obtained from Swedish national data registries. Annual incidence and prevalence of fungal disease was calculated based on epidemiological studies. Data on infections due to Cryptococcus sp., Mucorales, Histoplasma capsulatum, Coccidioides immitis and Pneumocystis jirovecii were retrieved from Karolinska University Laboratory and covers only 25% of Swedish population. In 2016, the population of Sweden was 9 995 153 (49.8% female). The overall burden of fungal infections was 1 713 385 (17 142/100 000). Superficial fungal infections affect 1 429 307 people (1429/100 000) based on Global Burden of Disease 14.3% prevalence. Total serious fungal infection burden was 284 174 (2843/100 000) in 2016. Recurrent Candida vulvovaginitis is common; assuming a 6% prevalence in women. Prevalence of allergic bronchopulmonary aspergillosis and severe asthma with fungal sensitisation were estimated to be 20 095 and 26 387, respectively. Similarly, chronic pulmonary aspergillosis was estimated to affect 490 patients after tuberculosis, sarcoidosis and other conditions. Candidemia incidence was estimated to be 500 in 2016 (4.7/100 000) and invasive aspergillosis 295 (3.0/100 000). In Stockholm area, Mucorales were reported in three patients in 2015, while Cryptococcus spp. were reported in two patients. In 2016, there were 297 patients PCR positive for P jirovecii. The present study shows that the overall burden of fungal infections in Sweden is high and affects 17% of the population. The morbidity, mortality and the healthcare-related costs due to fungal infections warrant further studies.


Assuntos
Efeitos Psicossociais da Doença , Micoses/epidemiologia , Adolescente , Idoso , Criança , Pré-Escolar , Dermatomicoses/epidemiologia , Dermatomicoses/microbiologia , Feminino , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Micoses/microbiologia , Prevalência , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/microbiologia , Suécia/epidemiologia , Vulvovaginite/epidemiologia , Vulvovaginite/microbiologia
4.
BMC Infect Dis ; 19(1): 658, 2019 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-31337356

RESUMO

BACKGROUND: Pneumocystis jirovecii pneumonia (PCP) is one of the most common HIV-related opportunistic infections. The diagnosis of PCP is based on analyses from respiratory tract specimens which may require the invasive procedure of a diagnostic bronchoscopy. The objective of this study was to evaluate the diagnostic potential of Pneumocystis jirovecii PCR in serum combined with the 1,3-ß-D-glucan (betaglucan) test for the diagnosis of PCP in HIV-infected patients. METHODS: This was a retrospective case-control study including serum samples from 26 HIV-infected patients with PCP collected within 5 days prior to the start of PCP treatment, 21 HIV-infected control subjects matched by blood CD4+ cell counts, and 18 blood donors. The serum samples were analyzed for Pneumocystis jirovecii PCR and betaglucan. The reference standard for PCP was based on previously described microbiological and clinical criteria. RESULTS: All patients with PCP had detectabe Pneumocystis jirovecii DNA in serum yielding a sensitivity for the Pneumocystis jirovecii PCR assay in serum of 100%. All blood donors had negative Pneumocystis PCR in serum. The specificity when testing HIV-infected patients was 71%, but with a PCR Cycle threshold (Ct) value of 34 as cut-off the specificity was 90%. At a putative pretest probaility of 20%, the negative and positive predictive value for the Pneumocystis PCR assay in serum was 0.99 and 0.71, respectively. Betaglucan with cut-off level 200 pg/ml combined with a positive Pneumocystis jirovecii PCR result had sensitivity and specificity of 92 and 90%, respectively. The concentration of Pneumocystis jirovecii DNA in serum samples, expressed by the PCR Ct values, correlated inversely to the betaglucan levels in serum. CONCLUSION: In this case-control study including 70% of all HIV-infected patients with PCP treated at Sahlgrenska University Hospital during a time period of 13 years, Pneumocystis PCR analysis on serum samples had a very high sensitivity and negative predictive value for the diagnosis of PCP in HIV-infected patients. A serum-based diagnostic procedure either based on Pneumocystis jirovecii PCR alone or in combination with betaglucan analysis may thus be feasible and would facilitate the care of HIV-infected patients with suspected PCP.


Assuntos
Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Pneumocystis carinii/genética , Pneumonia por Pneumocystis/microbiologia , beta-Glucanas/sangue , Adolescente , Adulto , Idoso , Doadores de Sangue , Estudos de Casos e Controles , DNA Fúngico/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pneumocystis carinii/patogenicidade , Pneumonia por Pneumocystis/sangue , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Sensibilidade e Especificidade
5.
Sci Rep ; 9(1): 3838, 2019 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-30846717

RESUMO

Candida albicans is the most frequently isolated fungal species in hospital settings worldwide. However, non-albicans Candida species with decreased susceptibility to antifungals have emerged as an important cause of fungemia. The aims of this study were to determine the species distribution of fungi isolated from the blood samples of patients at a Swedish University Hospital and to define the in vitro susceptibilities of these isolates to nine antifungal agents. In total, 233 yeast isolates from 143 patients were included in this study. Antifungal susceptibility testing was performed using broth dilution Sensititre YeastOne panels, which comprised amphotericin B, 5-flucytosine, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, micafungin, and caspofungin. The most common species in all age groups was C. albicans (n = 93, 65%), followed by C. glabrata (n = 27, 19%) and C. parapsilosis (n = 15, 10%). C. glabrata was mostly found in elderly individuals, while C. parapsilosis was found mainly in young children (p = 0.008). Antifungal resistance was low in the Candida species, except for reduced susceptibility to fluconazole among C. glabrata strains. C. albicans is the most frequent colonizer of Swedish patients. In general antifungal resistance is uncommon in Candida species. Nevertheless, reduced susceptibilities to fluconazole and echinocandins were found in C. glabrata and C. parapsilosis, respectively.


Assuntos
Antifúngicos/uso terapêutico , Candida/efeitos dos fármacos , Candidemia/microbiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anfotericina B/uso terapêutico , Anidulafungina/uso terapêutico , Candida/isolamento & purificação , Candida albicans/efeitos dos fármacos , Candida albicans/isolamento & purificação , Candida glabrata/efeitos dos fármacos , Candida glabrata/isolamento & purificação , Candida parapsilosis/efeitos dos fármacos , Candida parapsilosis/isolamento & purificação , Candidemia/tratamento farmacológico , Caspofungina/uso terapêutico , Criança , Pré-Escolar , Feminino , Fluconazol/uso terapêutico , Flucitosina/uso terapêutico , Humanos , Itraconazol/uso terapêutico , Masculino , Micafungina/uso terapêutico , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Triazóis/uso terapêutico , Voriconazol/uso terapêutico , Adulto Jovem
6.
Mycoses ; 61(10): 777-785, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29920785

RESUMO

OBJECTIVES: To identify the epidemiology and antifungal susceptibilities of Candida spp. among blood culture isolates to identify the epidemiology and antifungal susceptibilities of Candida spp. among blood culture isolates in Sweden. METHODS: The study was a retrospective, observational nationwide laboratory-based surveillance for fungaemia and fungal meningitis and was conducted from September 2015 to August 2016. RESULTS: In total, 488 Candida blood culture isolates were obtained from 471 patients (58% males). Compared to our previous study, the incidence of candidaemia has increased from 4.2/100 000 (2005-2006) to 4.7/100 000 population/year (2015-2016). The three most common Candida spp. isolated from blood cultures were Candida albicans (54.7%), Candida glabrata (19.7%) and species in the Candida parapsilosis complex (9.4%). Candida resistance to fluconazole was 2% in C. albicans and between 0% and 100%, in non-albicans species other than C. glabrata and C. krusei. Resistance to voriconazole was rare, except for C. glabrata, C. krusei and C. tropicalis. Resistance to anidulafungin was 3.8% while no Candida isolate was resistant to amphotericin B. CONCLUSIONS: We report an overall increase in candidaemia but a minor decrease of C. albicans while C. glabrata and C. parapsilosis remain constant over this 10-year period.


Assuntos
Candida/classificação , Candida/isolamento & purificação , Candidemia/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Candidemia/etiologia , Criança , Pré-Escolar , Farmacorresistência Fúngica , Feminino , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Meningite Fúngica/epidemiologia , Meningite Fúngica/etiologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Estudos Retrospectivos , Suécia/epidemiologia , Adulto Jovem
8.
Mycoses ; 61(9): 623-632, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29577474

RESUMO

We prospectively evaluated a combination of fungal biomarkers in adult haematology patients with focus on their clinical utility at different time points during the course of infection. In total, 135 patients were monitored once to twice weekly for serum (1-3)-ß-d-glucan (BG), galactomannan (GM), bis-methyl-gliotoxin and urinary d-arabinitol/l-arabinitol ratio. In all, 13 cases with proven or probable invasive fungal disease (IFD) were identified. The sensitivity of BG and GM at the time of diagnosis (TOD) was low, but within 2 weeks from the TOD the sensitivity of BG was 92%. BG >800 pg/mL was highly specific for IFD. At a pre-test probability of 12%, both BG and GM had negative predictive values (NPV) >0.9 but low positive predictive values (PPV). In a subgroup analysis of patients with clinically suspected IFD (pre-test probability of 35%), the NPV was lower, but the PPV for BG was 0.86 at cut-off 160 pg/mL. Among IFD patients, 91% had patterns of consecutively positive and increasing BG levels. Bis-methyl-gliotoxin was undetectable in 15 patients with proven, probable and possible IA. To conclude, BG was the superior fungal marker for IFD diagnosis. Quantification above the limit of detection and graphical evaluation of the pattern of dynamics are warranted in the interpretation of BG results.


Assuntos
Biomarcadores/análise , Testes Diagnósticos de Rotina/métodos , Doenças Hematológicas/complicações , Infecções Fúngicas Invasivas/diagnóstico , Adolescente , Adulto , Idoso , Feminino , Gliotoxina/análogos & derivados , Gliotoxina/sangue , Humanos , Masculino , Mananas/sangue , Pessoa de Meia-Idade , Estudos Prospectivos , Sensibilidade e Especificidade , Soro/química , Álcoois Açúcares/urina , Urinálise , Adulto Jovem , beta-Glucanas/sangue
9.
Med Mycol ; 56(6): 695-702, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-29228273

RESUMO

Detection of species of Exophiala and Scedosporium in the respiratory tracts of cystic fibrosis (CF) patients remains controversial because of highly variable results. The results of our study suggested a significantly higher prevalence and more complex colonization than previously estimated. Approximately 17% (27/162) of clinical sputum samples were found to be positive for Exophiala dermatitidis and 30% (49/162) were positive for Scedosporium apiospermum / S. boydii species complex determined by reverse line blot (RLB) hybridization. In contrast, only 14.2% (23/162) and 1.2% (2/162) of clinical sputa were positive for E. dermatitidis and S. apiospermum / S. boydii species complex when tested by culture, respectively. Molecular detection methods, such as loop-mediated isothermal amplification (LAMP) or reverse line blot (RLB) hybridization, have the potential to become powerful alternatives to selective culture, providing a more realistic understanding on the prevalence of E. dermatitidis and S. apiospermum / S. boydii species complex in the respiratory tract of CF patients.


Assuntos
Fibrose Cística/complicações , Exophiala/isolamento & purificação , Micoses/complicações , Micoses/diagnóstico , Scedosporium/isolamento & purificação , Escarro/microbiologia , Exophiala/genética , Humanos , Técnicas de Amplificação de Ácido Nucleico , Reprodutibilidade dos Testes , Sistema Respiratório/microbiologia , Scedosporium/genética , Sensibilidade e Especificidade , Tubulina (Proteína)/genética
10.
Mycopathologia ; 183(1): 7-19, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29098487

RESUMO

The achievement of a better life for cystic fibrosis (CF) patients is mainly caused by a better management and infection control over the last three decades. Herein, we want to summarize the cornerstones for an effective management of CF patients and to give an overview of the knowledge about the fungal epidemiology in this clinical context in Europe. Data from a retrospective analysis encompassing 66,616 samples from 3235 CF patients followed-up in 9 CF centers from different European countries are shown.


Assuntos
Fibrose Cística/complicações , Gerenciamento Clínico , Pneumopatias Fúngicas/diagnóstico , Pneumopatias Fúngicas/tratamento farmacológico , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Europa (Continente)/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Pneumopatias Fúngicas/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Adulto Jovem
11.
Mol Pharm ; 13(10): 3595-3600, 2016 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-27576445

RESUMO

Novel antifungals are in high demand due to the challenges associated with resistant, persistent, and systemic fungal infections. Synthetic mimics of antimicrobial peptides are emerging as a promising class of compounds for antifungal treatment. In the current study, five synthetic cationic antimicrobial tripeptides were evaluated as antifungal therapeutics against 24 pathogenic strains of fungi. Three of the peptides displayed strong general antifungal properties at low micromolar inhibitory concentrations. The most promising peptide, compound 5, was selected and evaluated as an antifungal remedy for Candida albicans candidiasis in a human skin model and for the treatment of Trichophyton rubrum induced onychomycosis in an infected human nail model. Compound 5 was shown to display antifungal properties and a rapid mode of action superior to those of both the commercial comparators Loceryl and Lamisil. Compound 5 was also active against a clinical isolate of Candida albicans with acquired fluconazole resistance.


Assuntos
Antifúngicos/uso terapêutico , Peptídeos Catiônicos Antimicrobianos/farmacologia , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Candidíase/tratamento farmacológico , Morfolinas/uso terapêutico , Naftalenos/uso terapêutico , Onicomicose/tratamento farmacológico , Peptídeos Catiônicos Antimicrobianos/farmacocinética , Candida albicans/efeitos dos fármacos , Candida albicans/patogenicidade , Humanos , Testes de Sensibilidade Microbiana , Dermatopatias/tratamento farmacológico , Dermatopatias/microbiologia , Terbinafina
12.
Jundishapur J Microbiol ; 8(10): e22929, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26587207

RESUMO

BACKGROUND: Blood stream infections (BSIs) are major causes of morbidity and mortality in burn patients. Microorganisms responsible for BSI are generally bacteria; however, Candida spp. are the infection agents in as many as 8% of all cases. Burn wound colonization and infections are generally the first steps to systemic infection. Candidemia in burn patients has been associated with high mortality and a prolonged hospital stay. OBJECTIVES: Candidemia in burn patients has been defined as a preterminal event, leading to high morbidity and mortality rates among these patients. The aim of this study was to establish the incidence of candidemia in burn patients in Iran. PATIENTS AND METHODS: We consecutively collected 405 blood samples from 113 burn patients. The yeast isolates were identified to the species level using conventional procedures. In vitro antifungal susceptibility of the Candida isolates to amphotericin B, fluconazole, voriconazole and caspofungin was performed using the Etest. RESULTS: Twenty-seven samples (6.7%) of the blood cultures from 13 patients (12%) were positive for Candida species. Candida parapsilosis (38%) and C. tropicalis (38%) were the most commonly found Candida species, followed by C. albicans (15%) and C. guilliermondii (15%) in the patients. The incidence of candidemia was significantly correlated with increased duration of hospitalization, increased time of stay in the intensive care unit, and higher mortality. The antifungal susceptibility tests demonstrated that amphotericin B and voriconazole had the lowest minimum inhibitory concentrations (MICs) against Candida spp. CONCLUSIONS: Non-albicans Candida should be considered as significant pathogens in burned patients with candidemia.

13.
Med Mycol ; 53(7): 725-35, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26162476

RESUMO

We conducted a retrospective study to evaluate the usefulness of immunoglobulin G (IgG) subclasses against Candida cell wall fragments (CW) and phosphopeptidomannan (PPM) for the diagnosis of invasive candidiasis (IC). We analyzed 54 patients with IC (n = 19), Candida heavy colonization (HC; n = 16), and controls (no IC or HC, n = 19).In nonneutropenic patients (n = 47), the sensitivity and specificity values of IgG1 anti-CW and IgG2 anti-PPM in IC were 88%, 59%, and 88%, 94%, respectively. The areas under the receiver operating characteristic curves were 0.69 (0.51-0.88) and 0.901 (0.78-1.02), respectively. IgG1 mean values (arbitrary units) and 95% confidence interval were 46 (20-71), 42 (-0.38 to 84) and 20 (8.3-32) in IC, HC, and in controls, respectively, and discriminated IC but not HC from controls (P = .032, and P = .77, respectively). IgG2 mean values were 26 (9.2-42), 19 (4.4-33), and 3.2 (0.28-6.6) in IC, HC, and in controls, respectively, and discriminated both IC and HC from controls (P < .0001 and P = .035, respectively) but did not separate IC from HC (P = .2). IgG2 showed positivity as early as one day after the IC diagnosis. Antibodies were detected in only two out of a total of seven neutropenic patients.For both IC and HC patients, the diagnostic performance of IgG2 anti-PPM was better than the one of IgG1 anti-CW. In nonneutropenic patients, IgG2 anti-PPM accurately identified not only IC patients but also HC patients at high risk for IC. This marker may help clinicians in the initiation of early preemptive therapy.


Assuntos
Anticorpos Antifúngicos/imunologia , Antígenos de Fungos/imunologia , Candida/imunologia , Candidíase Invasiva/diagnóstico , Parede Celular/imunologia , Imunoglobulina G/sangue , Mananas/imunologia , Fosfopeptídeos/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Estudos Retrospectivos , Sensibilidade e Especificidade , Adulto Jovem
14.
FEMS Microbiol Lett ; 362(1): 1-6, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25790495

RESUMO

Conventional mycological identifications based on the recognition of morphological characteristics can be problematic. A relatively new methodology applicable for the identification of microorganisms is based on the exploitation of taxon- specific mass patterns recorded from abundant cell proteins directly from whole-cell preparations, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This study reports the application of MALDI-TOF MS for the differentiation and identifications of black yeasts, isolated from the respiratory tracts of patients with cystic fibrosis (CF). Initial phenotypic and DNA sequence-based analyses identified these isolates to be Exophiala dermatitidis. The type strains of E. dermatitidis (CBS 207.35(T)) and other species of Exophiala were included in the MALDI-TOF MS analyses to establish the references for comparing the mass spectra of the clinical isolates of Exophiala. MALDI-TOF MS analyses exhibited extremely close relationships among the clinical isolates and with the spectra generated from the type strain of E. dermatitidis. The relationships observed between the E. dermatitidis strains from the MALDI-TOF MS profiling analyses were supported by DNA sequence-based analyses of the rRNA ITS1 and ITS2 regions. These data demonstrated the applicability of MALDI-TOF MS as a reliable, rapid and cost-effective method for the identification of isolates of E. dermatitidis and other clinically relevant fungi and yeasts that typically are difficult to identify by conventional methods.


Assuntos
Exophiala/classificação , Exophiala/isolamento & purificação , Micoses/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Análise por Conglomerados , Fibrose Cística/complicações , DNA Fúngico/química , DNA Fúngico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Exophiala/química , Humanos , Filogenia , Análise de Sequência de DNA
15.
J Cyst Fibros ; 13(4): 391-9, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24439251

RESUMO

BACKGROUND: The clinical importance of airway colonisation by the fungus Exophiala dermatitidis in patients with cystic fibrosis (CF) is unclear. We have previously shown that E. dermatitidis frequently colonises the airways of patients with CF. The aims of the present study were to determine whether patients who are colonised by E. dermatitidis have detectable fungal antigens in the circulation, develop anti-fungal antibodies, and show signs of inflammation and impaired respiratory function. METHODS: We collected sputum and serum samples consecutively from 98 sputum-producing patients with CF aged more than 12 years. The serum samples were subjected to bacterial and fungal culturing and analyses for fungal antigens and inflammatory factors. RESULTS: E. dermatitidis was recovered from 17 (17%) patients, the same isolation rate as for Aspergillus fumigatus. There were no difference regarding the levels of ß-glucan in the sera from E. dermatitidis culture-positive and culture-negative patients with CF. Serological analysis revealed significantly higher levels of IgG antibodies to E. dermatitidis cell wall fragments in the E. dermatitidis culture-positive patients. Patients with higher level of E. dermatitidis IgG antibodies were more often colonised with non-tuberculous Mycobacteria, and less often with Staphylococcus aureus. The increased levels of IgG antibodies directed against E. dermatitidis were positively associated with higher white blood cell counts, increased erythrocyte sedimentation rate, pancreatic insufficiency, intravenous antibiotic treatment, and they were negatively associated with respiratory function (FEV1 % predicted). Overall, 4/17 Exophiala-positive patients were diagnosed as having symptomatic infection with E. dermatitidis and were treated with broad-spectrum azoles. CONCLUSION: E. dermatitidis triggers antibody production and may cause significant airway infection in patients with cystic fibrosis.


Assuntos
Anticorpos Anti-Idiotípicos/imunologia , Anticorpos Antifúngicos/imunologia , Fibrose Cística/imunologia , Exophiala/imunologia , Imunoglobulina G/imunologia , Feoifomicose/imunologia , Adolescente , Adulto , Antígenos de Fungos/sangue , Antígenos de Fungos/imunologia , Criança , Estudos Transversais , Fibrose Cística/sangue , Fibrose Cística/complicações , Exophiala/isolamento & purificação , Feminino , Seguimentos , Humanos , Técnicas Imunoenzimáticas , Inflamação/imunologia , Inflamação/patologia , Masculino , Feoifomicose/complicações , Feoifomicose/microbiologia , Estudos Retrospectivos , Escarro/imunologia , Escarro/microbiologia , Adulto Jovem
16.
Mycopathologia ; 176(3-4): 237-41, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23948965

RESUMO

The laboratory diagnosis of dermatophytosis is usually based on direct microscopic examination and culturing of clinical specimens. A commercial polymerase chain reaction kit (Dermatophyte PCR) has had favorable results when used for detection of dermatophytes and identification of Trichophyton rubrum in nail specimens. This study investigated the efficacy of the Dermatophyte PCR kit for detecting dermatophytosis in 191 hair or skin specimens from patients with suspected dermatophytosis. PCR was positive for 37 % of samples, whereas 31 and 39 % of the specimens were positive by culturing and direct microscopy, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value for PCR analysis were 83, 84, 71, and 91 %, respectively. The sensitivity of the PCR test was higher in specimens obtained from skin (88 %) than in those obtained from hair (58 %), while the specificity remained almost the same (84 and 86 % for skin and hair, respectively). Our results show that the Dermatophyte PCR kit is a promising diagnostic tool for detection of dermatophytosis in skin samples, providing clinicians with a rapid diagnosis.


Assuntos
Arthrodermataceae/isolamento & purificação , Técnicas Microbiológicas/métodos , Técnicas de Diagnóstico Molecular/métodos , Pele/microbiologia , Tinha/diagnóstico , Tinha/microbiologia , Cabelo/microbiologia , Humanos , Microscopia , Reação em Cadeia da Polimerase/métodos , Valor Preditivo dos Testes , Sensibilidade e Especificidade
17.
Clin Vaccine Immunol ; 18(6): 969-77, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21508164

RESUMO

Whereas patients with multiple myeloma (MM) have a well-documented susceptibility to infections, this has been less studied in other B-cell disorders, such as Waldenstrom's macroglobulinemia (WM) and monoclonal gammopathy of undetermined significance (MGUS). We investigated the humoral immunity to 24 different pathogens in elderly patients with MM (n = 25), WM (n = 16), and MGUS (n = 18) and in age-matched controls (n = 20). Antibody titers against pneumococci, staphylococcal alpha-toxin, tetanus and diphtheria toxoids, and varicella, mumps, and rubella viruses were most depressed in MM patients, next to lowest in WM and MGUS patients, and highest in the controls. In contrast, levels of antibodies specific for staphylococcal teichoic acid, Moraxella catarrhalis, candida, aspergillus, and measles virus were similarly decreased in MM and MGUS patients. Comparable titers in all study groups were seen against Haemophilus influenzae type b (Hib), borrelia, toxoplasma, and members of the herpesvirus family. Finally, a uniform lack of antibodies was noted against Streptococcus pyogenes, salmonella, yersinia, brucella, francisella, and herpes simplex virus type 2. To conclude, although MM patients displayed the most depressed humoral immunity, significantly decreased antibody levels were also evident in patients with WM and MGUS, particularly against Staphylococcus aureus, pneumococci, and varicella. Conversely, immunity was retained for Hib and certain herpesviruses in all study groups.


Assuntos
Infecções Bacterianas/imunologia , Candidíase/imunologia , Gamopatia Monoclonal de Significância Indeterminada/imunologia , Mieloma Múltiplo/imunologia , Toxoplasmose/imunologia , Viroses/imunologia , Macroglobulinemia de Waldenstrom/imunologia , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antibacterianos/sangue , Anticorpos Antifúngicos/sangue , Anticorpos Antiprotozoários/sangue , Anticorpos Antivirais/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Gamopatia Monoclonal de Significância Indeterminada/complicações , Mieloma Múltiplo/complicações , Macroglobulinemia de Waldenstrom/complicações
18.
J Clin Microbiol ; 49(3): 1004-9, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21209163

RESUMO

The black-pigmented fungus Exophiala dermatitidis is considered to be a harmless colonizer of the airways of cystic fibrosis (CF) patients. The aim of this study was to establish the recovery rate of E. dermatitidis in respiratory specimens from CF patients, transplant recipients, and subjects with other respiratory disorders in Sweden. Second, we wished to determine if particular clinical traits were associated with E. dermatitidis colonization of the airways and the antifungal susceptibility profiles of Exophiala strains. Sputum and bronchoalveolar lavage samples (n = 492) derived from 275 patients were investigated. E. dermatitidis was isolated in respiratory specimens from 19% (18/97) of the CF patients but in none of the other patient categories. All isolates were recovered after 6 to 25 days of incubation on erythritol-chloramphenicol agar (ECA) medium. Morphological and genetic analyses confirmed species identity. Pancreatic insufficiency was positively associated with the presence of E. dermatitidis in sputum samples (P = 0.0198). Antifungal susceptibility tests demonstrated that voriconazole and posaconazole had the lowest MICs against E. dermatitidis. In conclusion, E. dermatitidis is a frequent colonizer of the respiratory tract in CF patients in Sweden and appears to be associated with more advanced disease. Whether E. dermatitidis is pathogenic remains to be elucidated.


Assuntos
Portador Sadio/microbiologia , Fibrose Cística/complicações , Insuficiência Pancreática Exócrina/complicações , Exophiala/isolamento & purificação , Infecções Respiratórias/microbiologia , Adolescente , Adulto , Idoso , Antifúngicos/farmacologia , Líquido da Lavagem Broncoalveolar/microbiologia , Exophiala/classificação , Exophiala/efeitos dos fármacos , Exophiala/genética , Feminino , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Micoses/microbiologia , Prevalência , Escarro/microbiologia , Suécia , Adulto Jovem
19.
Med Mycol ; 48(7): 1005-8, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20370367

RESUMO

The performance of a new commercially available duplex PCR, which combines pan-dermatophyte PCR with a Trichophyton rubrum-specific PCR, was evaluated. This Dermatophyte PCR kit, which requires one day for laboratory diagnosis, was compared with the conventional methods of microscopy and culture that necessitate up to 4 weeks for final diagnosis of dermatophytosis. We studied 177 nail samples from patients with suspected onychomycosis by fluorescence microscopy (blankophore), cultures and the Dermatophyte PCR kit. More samples were positive by PCR (78/177, 44%) than by culture (59/177, 34%). T. rubrum was present in 95% of all culture-positive nail specimens, which was confirmed by PCR in 55/56 specimens. The positive predictive value, negative predictive value, specificity and sensitivity of the duplex PCR was 93%, 87%, 94% and 85%, respectively, when confirmed by positive culture, microscopy or both. Due to its sensitivity, specificity and rapidity, we conclude that this PCR is an attractive method for routine investigation of nail dermatophytosis in a clinical setting.


Assuntos
Técnicas de Tipagem Micológica/métodos , Unhas/microbiologia , Onicomicose/diagnóstico , Reação em Cadeia da Polimerase/métodos , Kit de Reagentes para Diagnóstico , Tinha/diagnóstico , Trichophyton/isolamento & purificação , DNA Fúngico/genética , DNA Intergênico/genética , Diagnóstico Diferencial , Humanos , Sensibilidade e Especificidade , Tinha/microbiologia , Trichophyton/genética
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