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1.
J Mycol Med ; 30(2): 100970, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32334948

RESUMO

A survey of mycology laboratories for antifungal susceptibility testing (AFST) was undertaken in France in 2018, to better understand the difference in practices between the participating centers and to identify the difficulties they may encounter as well as eventual gaps with published standards and guidelines. The survey captured information from 45 mycology laboratories in France on how they perform AFST (number of strains tested, preferred method, technical and quality aspects, interpretation of the MIC values, reading and interpretation difficulties). Results indicated that 86% of respondents used Etest as AFST method, with a combination of one to seven antifungal agents tested. Most of the participating laboratories used similar technical parameters to perform their AFST method and a large majority used, as recommended, internal and external quality assessments. Almost all the participating mycology laboratories (98%) reported difficulties to interpret the MIC values, especially when no clinical breakpoints are available. The survey highlighted that the current AFST practices in France need homogenization, particularly for MIC reading and interpretation.


Assuntos
Antifúngicos/uso terapêutico , Laboratórios , Testes de Sensibilidade Microbiana , Micologia , Prática Profissional/estatística & dados numéricos , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão/métodos , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão/normas , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão/estatística & dados numéricos , Farmacorresistência Fúngica , França , História do Século XXI , Humanos , Laboratórios/normas , Laboratórios/estatística & dados numéricos , Ensaio de Proficiência Laboratorial/métodos , Ensaio de Proficiência Laboratorial/estatística & dados numéricos , Testes de Sensibilidade Microbiana/métodos , Testes de Sensibilidade Microbiana/normas , Testes de Sensibilidade Microbiana/estatística & dados numéricos , Micologia/história , Micologia/métodos , Micologia/normas , Micologia/estatística & dados numéricos , Prática Profissional/normas , Controle de Qualidade , Inquéritos e Questionários
2.
J Cyst Fibros ; 18(3): 364-367, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30638825

RESUMO

Azole-resistant Aspergillus fumigatus (ARAF) has been reported in the domestic environment of patients at risk for aspergillosis. Here, we assessed the mother's and father's homes of an 18-year-old cystic fibrosis patient harbouring chronic colonisation with H285Y CYP51A azole-resistant isolate, in order to explore the link between environmental exposure and ARAF infection. In one dwelling, a very high overall contamination level was found (710-7.240 CFU/m3), with a predominance of A. fumigatus (640-6.490 CFU/m3), and ARAF showing the TR34/L98H mutation was isolated. Mycological follow-up of the patient showed the persistence of H285Y isolates, but no acquisition of TR34/L98H isolates was observed. This could be due to the low proportion of TR34/L98H isolates (<3%), or the establishment of preventative measures and dwelling remediation taken after the environmental investigation. Our data underlines the value of an environmental assessment to establish preventative measures and limit the risk of A. fumigatus exposure and ARAF acquisition.


Assuntos
Poluição do Ar em Ambientes Fechados , Antifúngicos , Aspergilose Broncopulmonar Alérgica , Aspergillus fumigatus , Azóis/farmacologia , Fibrose Cística , Adolescente , Poluição do Ar em Ambientes Fechados/análise , Poluição do Ar em Ambientes Fechados/prevenção & controle , Antifúngicos/administração & dosagem , Antifúngicos/efeitos adversos , Antifúngicos/classificação , Aspergilose Broncopulmonar Alérgica/diagnóstico , Aspergilose Broncopulmonar Alérgica/etiologia , Aspergilose Broncopulmonar Alérgica/terapia , Aspergillus fumigatus/efeitos dos fármacos , Aspergillus fumigatus/isolamento & purificação , Fibrose Cística/microbiologia , Fibrose Cística/fisiopatologia , Fibrose Cística/terapia , Farmacorresistência Fúngica , Exposição Ambiental/análise , Exposição Ambiental/prevenção & controle , Humanos , Masculino , Resultado do Tratamento
3.
J Hosp Infect ; 99(1): 68-74, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29432820

RESUMO

BACKGROUND: Cutaneous mucormycoses, mainly due to Lichtheimia (Absidia), have occurred on several occasions in the Burn Unit of the University Hospital of Lille, France. AIM: To investigate the potential vector role of non-sterile bandages used to hold in place sterile gauze used for wound dressing. METHODS: Mycological analysis by conventional culture, Mucorales real-time polymerase chain reaction (qPCR), and Lichtheimia species-specific qPCR were performed on eight crepe and six elasticized bandages that were sampled on two independent occasions in March 2014 and July 2016. Characteristics of the seven Lichtheimia mucormycoses which occurred in burn patients between November 2013 and July 2016 were also collected to assess the epidemiological relationship between potentially contaminated bandages and clinical infections. FINDINGS: One Lichtheimia corymbifera strain was isolated from a crepe bandage by culture, and Lichtheimia spp. qPCR was positive in six out of eight crepe and four out of six elasticized bandages. Using species-specific qPCR, Lichtheimia ramosa, Lichtheimia ornata, and L. corymbifera were identified in six out of ten, five out of ten, and four out of ten bandages, respectively. In patients with mucormycosis, L. ramosa and L. ornata were present in five and two cases, respectively. CONCLUSION: Our data support the utility of Mucorales qPCR for epidemiological investigations, the potential role of these bandages in cutaneous mucormycoses in burn patients in our centre, and, consequently, the need for sterile bandages for the dressing of extensive wounds.


Assuntos
Bandagens/microbiologia , Queimaduras/complicações , Dermatomicoses/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Mucorales/isolamento & purificação , Mucormicose/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Adulto , Idoso , Dermatomicoses/microbiologia , Feminino , França , Hospitais Universitários , Humanos , Masculino , Pessoa de Meia-Idade , Mucorales/genética , Mucormicose/microbiologia
4.
Indoor Air ; 28(2): 298-306, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29082624

RESUMO

Azole-resistant Aspergillus fumigatus (ARAF) has been reported in patients with chronic obstructive pulmonary disease (COPD) but has not been specifically assessed so far. Here, we evaluated ARAF prevalence in azole-naïve COPD patients and their homes, and assessed whether CYP51A mutations were similar in clinical and environmental reservoirs. Sixty respiratory samples from 41 COPD patients with acute exacerbation and environmental samples from 36 of these patient's homes were prospectively collected. A. fumigatus was detected in respiratory samples from 11 of 41 patients (27%) and in 15 of 36 domiciles (42%). Cyp51A sequencing and selection on itraconazole medium of clinical (n = 68) and environmental (n = 48) isolates yielded ARAF detection in 1 of 11 A. fumigatus colonized patients with COPD (9%) and 2 of 15 A. fumigatus-positive patient's homes (13%). The clinical isolate had no CYP51A mutation. Two environmental isolates from two patients harbored TR34 /L98H mutation, and one had an H285Y mutation. Coexistence of different cyp51A genotypes and/or azole resistance profiles was detected in 3 of 8 respiratory and 2 of 10 environmental samples with more than one isolate, confirming the need for a systematic screening of all clinically relevant isolates. The high prevalence of ARAF in patients with COPD and their homes supports the need for further studies to assess the prevalence of azole resistance in patients with Aspergillus diseases in Northern France.


Assuntos
Poluição do Ar em Ambientes Fechados/análise , Antifúngicos/farmacologia , Aspergillus fumigatus/isolamento & purificação , Azóis/farmacologia , Doença Pulmonar Obstrutiva Crônica/microbiologia , Doença Aguda , Idoso , Aspergillus fumigatus/efeitos dos fármacos , Aspergillus fumigatus/genética , Contagem de Colônia Microbiana , Sistema Enzimático do Citocromo P-450/efeitos dos fármacos , Sistema Enzimático do Citocromo P-450/isolamento & purificação , Progressão da Doença , Farmacorresistência Fúngica/genética , Feminino , Proteínas Fúngicas/efeitos dos fármacos , Proteínas Fúngicas/isolamento & purificação , Genótipo , Habitação , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Prospectivos
5.
Rev Mal Respir ; 34(10): 1124-1137, 2017 Dec.
Artigo em Francês | MEDLINE | ID: mdl-29153274

RESUMO

INTRODUCTION: To study the impact of the constant and inevitable inhalation of moulds, it is necessary to sample, identify and count the spores. BACKGROUND: Environmental sampling methods can be separated into three categories: surface sampling is easy to perform but non quantitative, air sampling is easy to calibrate but provides time limited information, and dust sampling which is more representative of long term exposure to moulds. The sampling strategy depends on the objectives (evaluation of the risk of exposure for individuals; quantification of the household contamination; evaluation of the efficacy of remediation). The mould colonies obtained in culture are identified using microscopy, Maldi-TOF, and/or DNA sequencing. VIEWPOINTS: Electrostatic dust collectors are an alternative to older methods for identifying and quantifying household mould spores. They are easy to use and relatively cheap. Colony counting should be progressively replaced by quantitative real-time PCR, which is already validated, while waiting for more standardised high throughput sequencing methods for assessment of mould contamination without technical bias. CONCLUSION: Despite some technical recommendations for obtaining reliable and comparable results, the huge diversity of environmental moulds, the variable quantity of spores inhaled and the association with other allergens (mites, plants) make the evaluation of their impact on human health difficult. Hence there is a need for reliable and generally applicable quantitative methods.


Assuntos
Poluição do Ar em Ambientes Fechados/análise , Monitoramento Ambiental/métodos , Fungos/isolamento & purificação , Tipagem Molecular/métodos , Técnicas de Tipagem Micológica/métodos , Alérgenos/análise , Animais , Poeira/análise , Fungos/classificação , Fungos/genética , Humanos
6.
Eur J Clin Microbiol Infect Dis ; 35(1): 137-48, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26610340

RESUMO

Cryptosporidiosis is an important though underreported public health concern. Molecular tools might be helpful in improving its diagnosis. In this study, ZR Fecal DNA MiniPrep™ Kit (ZR) and NucliSens® easyMAG® (EM) were compared using four Cryptosporidium-seeded feces and 29 Cryptosporidium-positive stools. Thereafter, ZR was selected for prospective evaluation of Cryptosporidium detection by 18S rDNA and LAXER quantitative PCR (qPCR) in 69 stools from 56 patients after Cryptosporidium detection by glycerin, modified Ziehl-Neelsen (ZN) and auramine-phenol (AP) stainings. The combination of any of the two extraction methods with 18S qPCR yielded adequate detection of Cryptosporidium in seeded stools, but the ZR kit showed the best performance. All 29 Cryptosporidium-positive samples were positive with 18S qPCR, after both ZR and EM extraction. However, false-negative results were found with LAXER qPCR or nested PCR. Cryptosporidiosis was diagnosed in 7/56 patients. All the microscopic methods enabled the initial diagnosis, but Cryptosporidium was detected in 12, 13, and 14 samples from these seven patients after glycerin, ZN, and AP staining respectively. Among these samples, 14 and 12 were positive with 18S and LAXER qPCR respectively. In two patients, Cryptosporidium DNA loads were found to be correlated with clinical evolution. Although little known, glycerin is a sensitive method for the initial detection of Cryptosporidium. When combined with 18S qPCR, ZR extraction, which had not been evaluated so far for Cryptosporidium, was an accurate tool for detecting Cryptosporidium and estimating the oocyst shedding in the course of infection.


Assuntos
Criptosporidiose/diagnóstico , Cryptosporidium/isolamento & purificação , Microscopia/métodos , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase/métodos , Adolescente , Adulto , Criança , DNA de Protozoário/genética , DNA Ribossômico/genética , Reações Falso-Negativas , Feminino , Humanos , Masculino , RNA Ribossômico 18S/genética , Coloração e Rotulagem/métodos
7.
Med Mycol ; 53(7): 754-9, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26162469

RESUMO

Pneumocystis is mostly found in the alveolar spaces, but circulation of viable organisms also occurs and suggests that the detection of DNA in blood could be used as a noninvasive procedure to improve the diagnosis of Pneumocystis pneumonia (PcP). In order to determine the optimal compartment for Pneumocystis DNA detection, we used a rat model of PcP and tested the presence of Pneumocystis with a quantitative mtLSU targeting real-time PCR in four blood compartments: whole blood, clot, serum and Platelet-Rich-Plasma (PRP). All samples from 4 Pneumocystis-free control rats were negative. Pneumocystis was detected in 79, 64, 57, and 57% of samples from 14 PcP rats, respectively, but DNA release was not related to pulmonary loads. These data confirm the potential usefulness of Pneumocystis DNA detection in the blood for PcP diagnosis and suggest that whole blood could be the most appropriate compartment for Pneumocystis detection.


Assuntos
Sangue/microbiologia , DNA Fúngico/sangue , Pneumocystis carinii/isolamento & purificação , Pneumonia por Pneumocystis/diagnóstico , Pneumonia por Pneumocystis/microbiologia , Animais , DNA Fúngico/química , DNA Fúngico/genética , DNA Mitocondrial/genética , DNA Ribossômico/genética , Modelos Animais de Doenças , Pneumocystis carinii/genética , RNA Ribossômico/genética , Ratos Nus , Reação em Cadeia da Polimerase em Tempo Real
8.
New Microbes New Infect ; 6: 11-4, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26042187

RESUMO

Pneumocystis colonization may play a role in transmission and local inflammatory response. It was explored in patients with respiratory diseases in North Lebanon. Overall prevalence reached only 5.2% (95% CI 2.13-10.47) but it was higher (17.3%) in the subpopulation of patients with chronic obstructive pulmonary disease (COPD). COPD was the only factor associated with a significantly increased risk of colonization. mtLSU genotyping revealed predominance of genotype 2, identified in five patients (71.4%), including one patient who had co-infection with genotype 3. These first data in North Lebanon confirm Pneumocystis circulation among patients with respiratory diseases and the potential for transmission to immunocompromised patients.

10.
Eur J Clin Microbiol Infect Dis ; 28(3): 223-32, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18763000

RESUMO

In order to improve invasive pulmonary aspergillosis (IPA) diagnosis, a real-time polymerase chain reaction (PCR) assay detecting Aspergillus spp. was developed. Its detection limit reached 2-20 conidia. The retrospective evaluation on 64 bronchoalveolar lavage (BAL) fluids from 57 patients at risk for IPA, including 20 probable and five proven IPA patients, revealed a 88% or 38% sensitivity in direct examination (DE)/culture-positive or culture-negative BAL, respectively, whereas galactomannan (GM) sensitivity reached 88% or 58%, respectively. Influence on the Aspergillus-PCR yield of BAL fluid volume, cellular count and DNA content (evaluated by human beta-globin quantification) was assessed. Significantly higher beta-globin levels were detected in Aspergillus PCR-positive (median 5,112 pg/microl) than negative (median 1,332 pg/microl) BAL fluids, suggesting that the beta-globin level could reflect BAL yields and DNA extraction. Using beta-globin for the interpretation of fungal PCR could improve the negative predictive value of this test.


Assuntos
Aspergilose/diagnóstico , Líquido da Lavagem Broncoalveolar/microbiologia , Lavagem Broncoalveolar , Neoplasias Hematológicas/complicações , Reação em Cadeia da Polimerase/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Aspergillus/isolamento & purificação , DNA Fúngico/genética , Feminino , Galactose/análogos & derivados , Humanos , Masculino , Mananas/sangue , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Sensibilidade e Especificidade
11.
Ann Biol Clin (Paris) ; 65(3): 257-64, 2007.
Artigo em Francês | MEDLINE | ID: mdl-17502297

RESUMO

In addition to the serological systematic screening tests, kits to measure the avidity of toxoplasma IgG antibodies are currently available. Since high-avidity IgG toxoplasma antibodies have been shown to exclude recent infection, IgG avidity determination is especially useful in ruling out acute infection having occurred in the 3-4 prior months of pregnancy. We therefore compared the efficacy of two toxoplasma IgG avidity ELISA kits: SFRI (SFRI Laboratoire) and VIDAS Toxo-IgG avidity kit (bioMérieux). The agreement of the results from the 2 commercial assays were analysed using 55 serum samples, in terms of global mother-child Toxoplasma results and outcome, specially with light of the results of Toxoplasma antenatal, postnatal assays and of clinical follow up of children.


Assuntos
Anticorpos Antiprotozoários/imunologia , Afinidade de Anticorpos , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/imunologia , Complicações Infecciosas na Gravidez/sangue , Kit de Reagentes para Diagnóstico , Toxoplasma/imunologia , Toxoplasmose/sangue , Animais , Feminino , Humanos , Vigilância da População , Gravidez , Estudos Retrospectivos , Testes Sorológicos/métodos
12.
Ann Biol Clin (Paris) ; 63(1): 67-73, 2005.
Artigo em Francês | MEDLINE | ID: mdl-15689314

RESUMO

Congenital toxoplasmosis results from foetus contamination by Toxoplasma gondii during pregnancy. It is a frequent and severe condition calling for close monitoring of mothers at risk. During the last decades, numerous advances have been made specially in the antenatal diagnosis. The congenital toxoplasmosis diagnosis relies currently on PCR test of amniotic fluid, with a sensitivity of 80%. More recently, real-time quantitative PCR has been developed to improve toxoplasmosis diagnosis. We therefore compared the diagnosis value of quantitative real-time PCR with our conventional PCR-hybridization for the diagnosis of congenital toxoplasmosis.


Assuntos
Reação em Cadeia da Polimerase/métodos , Toxoplasma/isolamento & purificação , Toxoplasmose Congênita/diagnóstico , Líquido Amniótico/parasitologia , Animais , Sequência de Bases , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Feminino , Humanos , Dados de Sequência Molecular , Gravidez , Complicações Parasitárias na Gravidez , Diagnóstico Pré-Natal , Toxoplasma/genética , Toxoplasmose Congênita/transmissão
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