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2.
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
4.
Rev. argent. microbiol ; 51(3): 214-220, set. 2019. tab
Artigo em Inglês | LILACS | ID: biblio-1041827

RESUMO

Reference fungal cultures (RFCs) are essential for the internal quality control of laboratories. The production of these cultures requires standardized procedures (IRAM 14950:2016 and ISO 17034:2016 standards) carried out by a recognized and accredited laboratory. The aim of this work was to produce RFC in paper disks of autochthonous strains, characterized by two, homogeneous and stable reference methods traceable at species level. RFC were produced using 14 regional species (7 yeasts and 7 filamentous fungi) from the fungal culture collection (DMic). Paper disks were impregnated with a culture suspension, dried and packed. Homogeneity, viability, identity and purity were verified. Short-and long-term stability at different temperatures and storage times were studied. Characterization of each strain allowed to confirm its identity and to ensure its traceability at international level. Produced batches were homogeneous and stable at -20 ±5 °C for 30 months. This method of production was adequate to produce homogeneous and stable RFC with phenotypic and genotypic characteristics correctly defined and internationally traceable. Standardized procedures were developed for the production of certified RFC that could be transferred to other microorganisms. Providing RFC that represent regional strains allows laboratories to produce more reliable results with a favorable impact on medical diagnosis, the environment or the food industry.


Los cultivos microbianos de referencia (CR) son imprescindibles para el control de calidad interno de los laboratorios. Asegurar su producción requiere de procedimientos estandarizados (IRAM 14950:2016 e ISO 17034:2016) realizados en un laboratorio reconocido y acreditado. El objetivo de este estudio fue producir cultivos fúngicos de referencia en discos de papel, a partir de un panel de cultivos autóctonos caracterizados por dos métodos de referencia, trazables a nivel taxonómico de especie, homogéneos y estables. Se produjeron CR de 14 especies circulantes en Argentina (7 de levaduras y 7 de hongos miceliales), depositadas en la colección de hongos de interés médico (DMic). Los discos de papel fueron embebidos con una suspensión del cultivo por producir, secados y envasados. Se verificó la homogeneidad, viabilidad, identidad y pureza de cada lote. Se evaluó la estabilidad a corto y largo plazo a distintas temperaturas y tiempos de almacenamiento. La caracterización de cada CR nos permitió confirmar su identidad y asegurar su trazabilidad a nivel internacional. Los lotes producidos fueron homogéneos y estables durante 30 meses conservados a -20 ±5 °C. Este método resultó adecuado para producir CR homogéneos y estables, con características fenotípicas y genotípicas correctamente definidas y trazables a nivel internacional. Los procedimientos estandarizados desarrollados en este trabajo pueden ser transferidos para producir CR certificados de otros microorganismos. La provisión de CR que represente cepas regionales permite a los laboratorios producir resultados más confiables con un impacto favorable en el diagnóstico médico, los estudios ambientales y la industria alimenticia.


Assuntos
Bancos de Espécimes Biológicos , Fungos , Micologia/normas , Preservação Biológica/instrumentação , Preservação Biológica/métodos , Controle de Qualidade , Padrões de Referência , Leveduras , Meios de Cultura , Micologia/métodos
5.
Rev Argent Microbiol ; 51(3): 214-220, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30591317

RESUMO

Reference fungal cultures (RFCs) are essential for the internal quality control of laboratories. The production of these cultures requires standardized procedures (IRAM 14950:2016 and ISO 17034:2016 standards) carried out by a recognized and accredited laboratory. The aim of this work was to produce RFC in paper disks of autochthonous strains, characterized by two, homogeneous and stable reference methods traceable at species level. RFC were produced using 14 regional species (7 yeasts and 7 filamentous fungi) from the fungal culture collection (DMic). Paper disks were impregnated with a culture suspension, dried and packed. Homogeneity, viability, identity and purity were verified. Short- and long-term stability at different temperatures and storage times were studied. Characterization of each strain allowed to confirm its identity and to ensure its traceability at international level. Produced batches were homogeneous and stable at -20±5°C for 30 months. This method of production was adequate to produce homogeneous and stable RFC with phenotypic and genotypic characteristics correctly defined and internationally traceable. Standardized procedures were developed for the production of certified RFC that could be transferred to other microorganisms. Providing RFC that represent regional strains allows laboratories to produce more reliable results with a favorable impact on medical diagnosis, the environment or the food industry.


Assuntos
Bancos de Espécimes Biológicos , Fungos , Micologia/normas , Meios de Cultura , Micologia/métodos , Preservação Biológica/instrumentação , Preservação Biológica/métodos , Controle de Qualidade , Padrões de Referência , Leveduras
6.
Med Mycol ; 56(4): 416-425, 2018 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-29036605

RESUMO

An online survey of mycology laboratories in seven Asian countries was conducted to assess the status, competence, and services available. Country representatives from the Asia Fungal Working Group (AFWG) contacted as many laboratories performing mycology diagnosis as possible in their respective countries, requesting that the laboratory heads complete the online survey. In total, 241 laboratories responded, including 71 in China, 104 in India, 11 in Indonesia, 26 in the Philippines, four in Singapore, 18 in Taiwan, and seven in Thailand. Overall, 129/241 (53.5%) surveyed mycology laboratories operate as separate designated mycology laboratories, 75/241 (31.1%) conduct regular formal staff training, 103/241 (42.7%) are accredited, and 88/157 (56.1%) participate in external quality assurance scheme (EQAS) programs. Microscopy and culture methods are available in nearly all laboratories, although few perform DNA sequencing (37/219; 16.9%) or use matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS) (27/219; 12.3%) for isolate identification. Antifungal susceptibility testing is performed in 142/241 (58.9%) laboratories, mainly for yeasts. The most commonly performed nonculture diagnostic is cryptococcal antigen testing (66 laboratories), followed by galactomannan testing (55), polymerase chain reaction (PCR) diagnosis (37), and beta-D-glucan testing (24). Therapeutic drug monitoring is conducted in 21 laboratories. There is almost no access to advanced diagnostic tests, like galactomannan, ß-D-glucan, and PCR, in the surveyed laboratories in Indonesia, the Philippines, and Thailand. These results highlight the need for development of quality laboratories, accreditation and training of manpower in existing laboratories, and access to advanced non-culture-based diagnostic tests to facilitate the diagnosis of fungal infections in Asia.


Assuntos
Fungos/isolamento & purificação , Laboratórios/estatística & dados numéricos , Técnicas de Tipagem Micológica/estatística & dados numéricos , Micologia/estatística & dados numéricos , Micoses/diagnóstico , Ásia , Países em Desenvolvimento , Fungos/classificação , Humanos , Agências Internacionais , Laboratórios/normas , Técnicas de Tipagem Micológica/normas , Micologia/instrumentação , Micologia/normas , Micoses/microbiologia , Inquéritos e Questionários
7.
Rev. esp. quimioter ; 30(supl.1): 1-7, sept. 2017.
Artigo em Inglês | IBECS | ID: ibc-165928

RESUMO

Antimicrobial resistance in complex models of continuous infection is a current issue. The update 2017 course addresses about microbiological, epidemiological and clinical aspects useful for a current approach to infectious disease. During the last year, nosocomial pneumonia approach guides, recommendations for management of yeast and filamentous fungal infections, review papers on the empirical approach to peritonitis and extensive guidelines on stewardship have been published. HIV infection is being treated before and more intensively. The implementation of molecular biology, spectrometry and inmunology to traditional techniques of staining and culture achieve a better and faster microbiological diagnosis. Finally, the infection is increasingly integrated, assessing non-antibiotic aspects in the treatment (AU)


La resistencia a los antimicrobianos en modelos cada vez más complejos de infección continúa siendo actualidad. El curso de actualización de este año 2017 trata aspectos microbiológicos, epidemiológicos y clínicos útiles para un abordaje actual de la patología infecciosa. Durante el último año se han publicado guías de aproximación a la neumonía nosocomial, recomendaciones sobre el manejo de la infección fúngica por levaduras y filamentosos, documentos de revisión sobre el abordaje empírico de la peritonitis y una extensas guías sobre stewardship. En la infección por el VIH, cada vez se trata antes y más intensamente. La implementación de la biología molecular, la espectrometría y la inmunología a las técnicas tradicionales de tinción y cultivo consiguen un diagnóstico microbiológico mejor y más rápido. Por último, la infección se aborda de forma cada vez más integral, valorando aspectos no antibióticos en el tratamiento (AU)


Assuntos
Humanos , Doenças Transmissíveis/tratamento farmacológico , Doenças Transmissíveis/epidemiologia , Anti-Infecciosos/administração & dosagem , HIV , Bacteriologia/organização & administração , Bacteriologia/normas , Micologia/organização & administração , Micologia/normas
8.
Enferm Infecc Microbiol Clin ; 34 Suppl 3: 1-7, 2016 Jul.
Artigo em Espanhol | MEDLINE | ID: mdl-27474240

RESUMO

The External Quality Control Program of the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC) include controls for bacteriology, serology, mycology, parasitology, mycobacteria, virology and molecular microbiology. This article presents the most relevant conclusions and lessons from the 2014 controls. As a whole, the results obtained in 2014 confirm the excellent skill and good technical standards found in previous editions. However, erroneous results can be obtained in any laboratory and in clinically relevant determinations. Once again, the results of the SEIMC program highlighted the need to implement both internal and external controls in order to assure the maximal quality of the microbiological tests.


Assuntos
Doenças Transmissíveis/diagnóstico , Laboratórios/normas , Controle de Qualidade , Bacteriologia/normas , Micologia/normas , Padrões de Referência , Espanha , Virologia/normas
9.
Enferm. infecc. microbiol. clín. (Ed. impr.) ; 34(supl.3): 1-7, jul. 2016. tab
Artigo em Espanhol | IBECS | ID: ibc-170847

RESUMO

Se presenta el análisis anual de los resultados remitidos durante el año 2014 por los participantes inscritos en el Programa de Control de Calidad de la Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica (SEIMC), que incluye las áreas de bacteriología, serología, micología, parasitología, micobacteriología, virología y microbiología molecular. Los resultados obtenidos por los centros participantes resaltan, de nuevo, la adecuada capacitación de la gran mayoría de los laboratorios españoles de microbiología clínica, como ya venía sucediendo en los últimos años. Sin embargo, el programa muestra que es posible obtener resultados erróneos, incluso en determinaciones con importante trascendencia y en cualquier laboratorio. Una vez más, se destaca la importancia de complementar el control interno que cada laboratorio lleva a cabo con estudios de intercomparación externos, como los que ofrece el programa SEIMC


The External Quality Control Program of the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC) include controls for bacteriology, serology, mycology, parasitology, mycobacteria, virology and molecular microbiology. This article presents the most relevant conclusions and lessons from the 2014 controls. As a whole, the results obtained in 2014 confirm the excellent skill and good technical standards found in previous editions. However, erroneous results can be obtained in any laboratory and in clinically relevant determinations. Once again, the results of the SEIMC program highlighted the need to implement both internal and external controls in order to assure the maximal quality of the microbiological tests


Assuntos
Controle de Qualidade , Laboratórios/normas , Doenças Transmissíveis/diagnóstico , Virologia/normas , Bacteriologia/normas , Micologia/normas , Espanha , Padrões de Referência
10.
Braz. j. microbiol ; 47(2): 403-409, Apr.-June 2016. tab, graf
Artigo em Inglês | LILACS | ID: lil-780836

RESUMO

Abstract Considering the absence of standards for culture collections and more specifically for biological resource centers in the world, in addition to the absence of certified biological material in Brazil, this study aimed to evaluate a Fungal Collection from Fiocruz, as a producer of certified reference material and as Biological Resource Center (BRC). For this evaluation, a checklist based on the requirements of ABNT ISO GUIA34:2012 correlated with the ABNT NBR ISO/IEC17025:2005, was designed and applied. Complementing the implementation of the checklist, an internal audit was performed. An evaluation of this Collection as a BRC was also conducted following the requirements of the NIT-DICLA-061, the Brazilian internal standard from Inmetro, based on ABNT NBR ISO/IEC 17025:2005, ABNT ISO GUIA 34:2012 and OECD Best Practice Guidelines for BRCs. This was the first time that the NIT DICLA-061 was applied in a culture collection during an internal audit. The assessments enabled the proposal for the adequacy of this Collection to assure the implementation of the management system for their future accreditation by Inmetro as a certified reference material producer as well as its future accreditation as a Biological Resource Center according to the NIT-DICLA-061.


Assuntos
Preservação Biológica/normas , Fungos/classificação , Micologia/organização & administração , Controle de Qualidade , Brasil , Fungos/isolamento & purificação , Fungos/genética , Micologia/normas
11.
Braz J Microbiol ; 47(2): 403-9, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26991280

RESUMO

Considering the absence of standards for culture collections and more specifically for biological resource centers in the world, in addition to the absence of certified biological material in Brazil, this study aimed to evaluate a Fungal Collection from Fiocruz, as a producer of certified reference material and as Biological Resource Center (BRC). For this evaluation, a checklist based on the requirements of ABNT ISO GUIA34:2012 correlated with the ABNT NBR ISO/IEC17025:2005, was designed and applied. Complementing the implementation of the checklist, an internal audit was performed. An evaluation of this Collection as a BRC was also conducted following the requirements of the NIT-DICLA-061, the Brazilian internal standard from Inmetro, based on ABNT NBR ISO/IEC 17025:2005, ABNT ISO GUIA 34:2012 and OECD Best Practice Guidelines for BRCs. This was the first time that the NIT DICLA-061 was applied in a culture collection during an internal audit. The assessments enabled the proposal for the adequacy of this Collection to assure the implementation of the management system for their future accreditation by Inmetro as a certified reference material producer as well as its future accreditation as a Biological Resource Center according to the NIT-DICLA-061.


Assuntos
Fungos/classificação , Micologia/organização & administração , Preservação Biológica/normas , Brasil , Fungos/genética , Fungos/isolamento & purificação , Micologia/normas , Controle de Qualidade
12.
Med Mycol ; 54(4): 361-9, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26768372

RESUMO

Quantitative cerebrospinal fluid (CSF) cultures provide a measure of disease severity in cryptococcal meningitis. The fungal clearance rate by quantitative cultures has become a primary endpoint for phase II clinical trials. This study determined the inter-assay accuracy of three different quantitative culture methodologies. Among 91 participants with meningitis symptoms in Kampala, Uganda, during August-November 2013, 305 CSF samples were prospectively collected from patients at multiple time points during treatment. Samples were simultaneously cultured by three methods: (1) St. George's 100 mcl input volume of CSF with five 1:10 serial dilutions, (2) AIDS Clinical Trials Group (ACTG) method using 1000, 100, 10 mcl input volumes, and two 1:100 dilutions with 100 and 10 mcl input volume per dilution on seven agar plates; and (3) 10 mcl calibrated loop of undiluted and 1:100 diluted CSF (loop). Quantitative culture values did not statistically differ between St. George-ACTG methods (P= .09) but did for St. George-10 mcl loop (P< .001). Repeated measures pairwise correlation between any of the methods was high (r≥0.88). For detecting sterility, the ACTG-method had the highest negative predictive value of 97% (91% St. George, 60% loop), but the ACTG-method had occasional (∼10%) difficulties in quantification due to colony clumping. For CSF clearance rate, St. George-ACTG methods did not differ overall (mean -0.05 ± 0.07 log10CFU/ml/day;P= .14) on a group level; however, individual-level clearance varied. The St. George and ACTG quantitative CSF culture methods produced comparable but not identical results. Quantitative cultures can inform treatment management strategies.


Assuntos
Líquido Cefalorraquidiano/microbiologia , Meningite Criptocócica/líquido cefalorraquidiano , Meningite Criptocócica/microbiologia , Micologia/métodos , Micologia/normas , Infecções Oportunistas Relacionadas com a AIDS/líquido cefalorraquidiano , Infecções Oportunistas Relacionadas com a AIDS/tratamento farmacológico , Infecções Oportunistas Relacionadas com a AIDS/microbiologia , Adulto , Antifúngicos/uso terapêutico , Feminino , Humanos , Limite de Detecção , Masculino , Meningite Criptocócica/tratamento farmacológico , Estudos Prospectivos , Índice de Gravidade de Doença
13.
Hautarzt ; 67(1): 23-6, 2016 Jan.
Artigo em Alemão | MEDLINE | ID: mdl-26552920

RESUMO

The ability of recognizing various clinical manifestations of mucocutaneous mycosis, making a diagnosis, and establishing a treatment is part of a dermatologist's daily routine. However, due to the fact that clinical manifestations, laboratory diagnostics, and treatment are performed in one hand, laboratory findings are properly classified and interpreted. Since new binding guidelines of the German Medical Association on quality assurance measures in medical laboratory testing came into force, there is much concern among dermatologists of how to comply with these new regulations. It is the intention of the authors to help our readers to implement these new rules in order to make sure that mycological diagnostics continue to be part of a dermatologist's professional work.


Assuntos
Dermatomicoses/diagnóstico , Dermatomicoses/microbiologia , Técnicas de Tipagem Micológica/normas , Micologia/normas , Guias de Prática Clínica como Assunto , Garantia da Qualidade dos Cuidados de Saúde/normas , Técnicas de Laboratório Clínico/normas , Alemanha , Humanos
14.
Rev. iberoam. micol ; 31(3): 193-196, jul.-sept. 2014.
Artigo em Inglês | IBECS | ID: ibc-127630

RESUMO

Background. The basidiomycetous yeast Cryptococcus gattii is an emerging and primary pathogen. There is a lack of information about its environmental spread outside outbreak regions in Mediterranean Europe, North and South America. Environmental sampling for C. gattii and molecular characterization of the obtained isolates will provide an insight into the global spread of the various genotypes. Methods. Woody debris of native divi-divi (Caesalpinia coriaria) trees were sampled across Bonaire, Dutch Caribbean. Colonies suspected for Cryptococcus species were subjected to standard mycology investigations and identification by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Isolates identified as C. gattii were subjected to amplified fragment length polymorphism genotyping, mating-type analysis and multi-locus sequence typing. Results. Ten colonies of C. gattii were cultured from different trunk hollows of the same divi-divi tree. Molecular characterization showed that all isolates were genotype AFLP6/VGII and mating-type α. Multi-locus sequence typing revealed that all isolates were genetically indistinguishable from each other. Conclusions. C. gattii is present in the environment of Bonaire, which suggests that this yeast is likely to be present in the environment of other Caribbean islands (AU)


Antecedentes. La levadura Cryptococcus gattii es un basidiomiceto emergente y patógeno primario. Existe poca información acerca de su dispersión en medio ambiente fuera de las regiones con brotes descritos por esta levadura en la Europa mediterránea, Norte y Sur de América. Los muestreos del medio ambiente para la búsqueda de C. gattii y la caracterización molecular de los aislamientos obtenidos puede proveer de una visión global sobre la dispersión de varios de sus genotipos. Métodos. Se tomaron muestras de residuos de madera de árboles nativos divi-divi (Caesalpinia coriaria) en Bonaire, Caribe Holandés. Las colonias susceptibles de pertenecer a las especies de Cryptococcus se sometieron a un estudio micológico estándar e identificación por espectrometría de masas MALDI-TOF (Matrix-Assisted Laser Desorption Ionization-Time of Flight). Los aislamientos identificados como C. gattii se sometieron a genotipado mediante AFLP (Amplified Fragment Length Polymorphism), obtención del tipo sexual y MLST (Multi-locus Sequence Typing). Resultados. Se obtuvieron diez colonias de C. gattii en el cultivo de nuestras de diferentes agujeros de un mismo árbol divi-divi. La caracterización molecular mostró que todos los aislamientos eran genotipo AFLP6/VGII y tipo sexual α. El tipado mediante MLST reveló que todos los aislamientos eran genéticamente indistinguibles unos de otros. Conclusiones. C. gattii está presente en el medio ambiente de Bonaire, lo que sugiere que esta levadura podría estar presente en el ambiente de otras islas del Caribe (AU)


Assuntos
Humanos , Masculino , Feminino , Cryptococcus gattii/isolamento & purificação , Caesalpinia/microbiologia , Genótipo , Poluição Ambiental/análise , Poluição Ambiental/prevenção & controle , Técnicas de Genotipagem/métodos , Técnicas de Genotipagem , Micologia/métodos , Micologia/normas , Micologia/tendências
15.
Rev. iberoam. micol ; 30(2): 109-111, abr.-jun. 2013.
Artigo em Espanhol | IBECS | ID: ibc-112582

RESUMO

Antecedentes. El diagnóstico micológico de onicomicosis puede establecerse mediante el examen microscópico directo (KOH) de una muestra clínica, el cultivo y la adición de blanco de calcoflúor. Objetivos. Comparar el porcentaje de positividad y el grado de correlación entre la adición de KOH, el cultivo y la tinción con blanco de calcoflúor en el diagnóstico de onicomicosis. Métodos. Estudio descriptivo, transversal y comparativo. Se utilizaron muestras de fragmentos ungueales de los pies para la adición de KOH, cultivo y adición de blanco de calcoflúor para examen bajo fluorescencia. Se calculó el porcentaje de positividad de las diferentes técnicas y se determinó el grado de correlación entre ellas (Programa Epi Info v 3.4.3©). Resultados. La adición de KOH dio un resultado positivo en el 66,67% de los casos, el cultivo en el 33,33% y la tinción con blanco de calcoflúor en el 57,58%. El KOH y el blanco de calcoflúor se asociaron a un mayor porcentaje de positividad que el cultivo (p < 0,01 y p < 0,05, respectivamente). El grado de correlación entre el KOH y el blanco de calcoflúor fue excelente (κ = 0,8085; p < 0,0001), mientras que fue débil entre el KOH y el cultivo y entre el blanco de calcoflúor y el cultivo. Conclusiones. No se recomienda el uso sistemático de blanco de calcoflúor en los laboratorios porque no parece conferir beneficios adicionales cuando se compara con el KOH. Esto reviste especial importancia cuando los recursos son limitados(AU)


Background. Mycological diagnosis of onychomycosis can be performed by direct microcopy (KOH), cultures and calcofluor white. Aims. To compare the percentage of positivity and the degree of correlation of KOH, cultures and calcofluor white for the diagnosis of onychomycosis. Methods. Descriptive, transversal and comparative study. Samples of toenails with onychomycosis were used for KOH, cultures and calcofluor white under fluorescence. The percentage of positivity of the different techniques was calculated and the degree of correlation between them was determined (Epi Info v 3.4.3©). Results. KOH was positive in 66.67% of the cases, cultures in 33.33% and calcofluor white in 57.58%. KOH and calcofluor white had a higher percentage of positivity than culture (p<0.01 and p<0.05 respectively). The degree of correlation between KOH and calcofluor white was excellent (κ=0.8085; p<0.0001); however, the degree of correlation between KOH and culture and between calcofluor white and culture was poor. Conclusions. The use of calcofluor white is not recommended in routine laboratories because it does not seem to bring any additional benefits when comparing with KOH. This is especially important when funding is a great problem(AU)


Assuntos
Humanos , Masculino , Feminino , Onicomicose/complicações , Onicomicose/diagnóstico , /instrumentação , /tendências , Microscopia/métodos , Microscopia/normas , Microscopia , Antifúngicos/uso terapêutico , Onicomicose/microbiologia , Micologia/métodos , Micologia/organização & administração , Micologia/normas , Estudos Transversais/métodos , Estudos Transversais/tendências , Estudos Transversais
17.
Med Mycol ; 51(5): 545-54, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23336696

RESUMO

Invasive aspergillosis (IA) is a serious nosocomial infection caused by Aspergillus spp. which has a high mortality rate due to the fact, among other factors, that it is difficult to diagnose. Within the Aspergillus genus, A. fumigatus is the main species causing IA. We propose a virulence factor, the aspHS gene, as a novel target for the specific detection of A. fumigatus by quantitative real-time PCR (qPCR). This target gene encodes a haemolysin, which is overexpressed in vivo during infection. We have designed specific primers and hydrolysis (Taqman) probes for the detection of this target and a chimeric internal amplification control (IC), designed to detect false negative results due to PCR inhibition. This qPCR assay was tested with DNA extracted from a wide collection of microorganisms, tissues from infected mice, and human bronchoalveolar lavage (BAL) samples. Results showed that it, together with the DNA extraction method, could detect A. fumigatus with high specificity. Furthermore, it can distinguish between germinated (first step to the development of infection) and non-germinated conidia (not detected). Our data indicate that these techniques could be sufficiently sensitive and rapid to help clinicians establish an earlier diagnosis, but the presence of PCR inhibitors in clinical samples such as BAL fluids needs to be addressed.


Assuntos
Aspergilose/diagnóstico , Aspergillus fumigatus/isolamento & purificação , Proteínas Fúngicas/genética , Técnicas de Diagnóstico Molecular/métodos , Micologia/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Fatores de Virulência/genética , Animais , Aspergillus fumigatus/genética , Primers do DNA/genética , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Técnicas de Diagnóstico Molecular/normas , Micologia/normas , Sondas de Oligonucleotídeos/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Sensibilidade e Especificidade
18.
Rev. iberoam. micol ; 30(1): 9-13, ene. 2013.
Artigo em Espanhol | IBECS | ID: ibc-109125

RESUMO

Antecedentes. En todo el mundo se considera que el maíz es uno de los cereales más vulnerables a las micotoxinas. Dos de las asociadas con más frecuencia a este cereal son las fumonisinas y la ocratoxina A. Se sabe que A. niger produce ocratoxina A, que suele estar presente en el maíz. Sin embargo, recientemente, se ha descrito que A. niger puede producir fumonisinas, principalmente fumonisina B2. Objetivos. El objetivo del presente estudio fue aislar las cepas de Aspergillus niger a partir de muestras de maíz provenientes de la siega en tres regiones agrícolas portuguesas y detectar la producción de fumonisina B2 y ocratoxina A. Métodos. Se obtuvieron 95 muestras de maíz de la siega en estas regiones, se sembraron en placas y se aislaron todas las cepas de Aspergillus sección Nigri observadas. Se caracterizaron morfológicamente las cepas y se determinó la producción de micotoxinas mediante cromatografía líquida de alto rendimiento con detección de fluorescencia. Resultados. Se aislaron un total de 270 cepas de Aspergillus sección Nigri de 73 muestras (77% de las muestras). Se observó que alrededor del 14% de las cepas producían ocratoxina A, mientras que aproximadamente el 39% producían fumonisina B2. Conclusiones. No pudo identificarse una asociación entre la producción de estas dos micotoxinas y no pudimos extraer conclusiones sobre si la presencia de cepas de A. niger aumentará el riesgo de contaminación del maíz por fumonisinas, en especial por fumonisina B2(AU)


Background. Maize is considered one of the crops more susceptible to mycotoxins in the world. Two of the mycotoxins commonly associated with maize are fumonisins and ochratoxin A. Aspergillus niger is a known producer of ochratoxin A and is easily found in maize. Recently, however, A. niger has been reported to produce as well fumonisins, mainly fumonisin B2. Aims. The aim of this study was to isolate A. niger strains from maize samples collected in three Portuguese maize growing regions and to detect the production of both fumonisin B2 and ochratoxin A. Methods. Ninety five maize samples were collected, plated, and all observable Aspergillus section Nigri strains were isolated. Strains were morphologically characterized and mycotoxin production was determined by HPLC-FD. Results. Isolations resulted in a total of 270 strains of black Aspergillus from 73 samples (77% of the samples). About 14% of those strains were found to produce ochratoxin A and 39% of the strains were found to produce fumonisin B2. Conclusions. An association between the production of these two mycotoxins could not be found and no conclusions could be taken whether the presence of A. niger aggregate strains will increase the risk of maize contamination with fumonisins and more specifically with fumonisin B2(AU)


Assuntos
Zea mays/química , Zea mays/microbiologia , Zea mays/normas , Micotoxinas/análise , Micotoxinas/isolamento & purificação , Fumonisinas/isolamento & purificação , Aspergillus niger , Micologia/métodos , Micologia/normas , Micologia/tendências
19.
Rev. iberoam. micol ; 30(1): 39-46, ene. 2013.
Artigo em Espanhol | IBECS | ID: ibc-109130

RESUMO

Antecedentes. En 1984 se crearon los Grupos de Trabajo en Micología de Venezuela (GTMV), proporcionando un abordaje novedoso al estudio de las micosis, en especial las micosis endémicas. Objetivos. Conocer los aportes en el estudio sistemático de las micosis en Venezuela durante 26 años de labor de los GTMV. Métodos. Se realizó revisión de la casuística publicada por los GTMV en el Boletín Informativo Las Micosis en Venezuela desde 1984 hasta 2010. Resultados. Se obtuvieron 36.968 diagnósticos de micosis superficiales, 1.989 de profundas sistémicas y 822 de profundas localizadas. La dermatofitosis fue la patología superficial más frecuente, paracoccidioidomicosis e histoplasmosis las profundas sistémicas, y cromoblastomicosis la profunda localizada. Se realizó la distribución geográfica de los casos de micosis profundas, pudiendo delimitar las zonas endémicas. Discusión. Las micosis superficiales constituyen un problema de salud pública por su alta morbilidad y pueden ser responsables de epidemias en grupos de riesgo. La paracoccidioidomicosis y la histoplamosis se reportaron con mayor frecuencia, incluso antes de haberse conformado los GTMV. El número de cromoblastomicosis y esporotricosis en Venezuela supera a lo reportado en otros países. Los GTMV han contribuido al conocimiento de la incidencia y prevalencia las micosis en el país, además de su divulgación como problema de salud pública, siendo un aporte invaluable que debe mantenerse en el tiempo, tratando no solo de reportar la casuística anual, sino también detallar aspectos clínico-epidemiológicos que permitan realizar seguimiento de la evolución de estas patologías(AU)


Background. In 1984 the Venezuelan Work Groups in Mycology (VWGM) were created introducing an innovative approach to the study of the mycoses in Venezuela. Aim. To study the occurrence of the mycoses in Venezuela. Methods. Review the reported cases of mycoses by the newsletter Boletín Informativo Las Micosis en Venezuela (VWGM) from 1984 to 2010. Results. The data collected showed 36,968 reported cases of superficial mycoses, 1,989 of deep systemic cases, and 822 of localized mycoses. Pityriasis dermatophytosis was the most common superficial infection, and paracoccidioidomycosis and histoplasmosis the most frequent deep systemic infection. Chromoblastomycosis was the most frequently diagnosed subcutaneous infection. The data provided showed the distribution by geographical area for each of the fungal infections studied, which may help to establish the endemic areas. Discussion. Superficial mycosis is a public health problem due to its high morbidity and is probably responsible for some of the outbreaks in high-risk groups. Paracoccidioidomycosis and histoplasmosis were reported more often, which agrees with earlier reports prior to the formation of the VWGM. Cases of sporotrichosis and chromoblastomycosis in Venezuela can be considered unique due to the high number of cases. This study highlights the contribution of the VWGM to the behavior of the mycoses in Venezuela, its incidence, prevalence, and the recognition of these infections as a problem of public health importance. The VWGM should keep working in this endeavor, not only reporting new cases, but also unifying the clinical and epidemiological criteria, in order to properly monitor the evolving epidemiological changes reported in these types of infections(AU)


Assuntos
Humanos , Masculino , Feminino , Micologia/métodos , Micologia/organização & administração , Micologia/normas , Micoses/epidemiologia , Micoses/microbiologia , Paracoccidioidomicose/epidemiologia , Paracoccidioidomicose/microbiologia , Esporotricose/epidemiologia , Esporotricose/microbiologia , Venezuela , Micologia/estatística & dados numéricos , Micologia/tendências , Histoplasmose/epidemiologia , Histoplasmose/microbiologia , Cromoblastomicose/epidemiologia , Cromoblastomicose/microbiologia , Indicadores de Morbimortalidade
20.
J Mycol Med ; 22(1): 14-20, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23177809

RESUMO

OBJECTIVE OF THE STUDY: Study of the spatio-temporal fungal colonization in a new medical mycology laboratory. METHODS: A 17-month survey of airborne fungal contamination was conducted in a new medical mycology laboratory at a tertiary care university hospital. This survey was implemented at three different periods: before the new premises were occupied (period A), during the move into the new laboratory (period B) and after resumption of the mycological activities in these new premises (period C). RESULTS: During period A, the airborne fungal load ranged from 2.3 to 6 cfu/m(3). The most frequently recovered airborne fungi were Penicillium spp. (75 to 100%). During period B, a dramatic increase in Penicillium chrysogenum conidia was observed in the air of the new laboratory (40 to 160 cfu/m(3)). During period C, the fungal load ranged from 4.5 to 8.4 cfu/m(3). Penicillium was the most common genus identified in rooms of the laboratory where no filamentous fungi were handled, while Aspergillus was clearly the predominant genus (78%) in the room dedicated to the culture of filamentous fungi. CONCLUSIONS: We suggest that the specific fungal ecology in air of the room dedicated to the culture of filamentous fungi is due to the handling of a large number of medical strains of A. fumigatus.


Assuntos
Microbiologia do Ar , Fungos/crescimento & desenvolvimento , Laboratórios Hospitalares , Micologia , Aspergillus/crescimento & desenvolvimento , Aspergillus/isolamento & purificação , Contagem de Colônia Microbiana , Monitoramento Ambiental , Unidades Hospitalares , Humanos , Laboratórios Hospitalares/normas , Micologia/normas , Penicillium/crescimento & desenvolvimento , Penicillium/isolamento & purificação
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