PCR multiplex en la detección de Tinea unguium en pacientes de Caracas – Venezuela // PCR multiplex in the detection of Tinea unguiumin patients from Caracas – Venezuela
Resumen
Resumen
LaTinea unguium(T. unguium) es una micosis superficial producida por dermatofitos que infectan la lámina ungueal de manos y pies. Afecta a más del 10% de la población mundial y su prevalencia viene aumentando en los últimos años. Los métodos convencionales para su diagnóstico, basados en el examen directo y el cultivo micológico, presentan baja especificidad y sensibilidad respectivamente. El reciente avance de técnicas de secuenciación del ADN, ha permitido desarrollar varios métodos moleculares para la identificación directa y más precisa de dermatofitos a partir de muestras clínicas, con resultados rápidos en 24-48 horas. El objetivo de este trabajo fue comparar la efectividad de la PCR multiplex con los métodos tradicionales para el diagnóstico de dermatofitos. Se evaluaron un total de 49 pacientes con diagnóstico clínico de T. unguiumy 10 muestras de individuos sanos. Se aplicaron las técnicas convencionales y la PCR multiplex para detectar secuencias comunes de los dermatofitos y secuencias específicas para Trichophyton (T.) rubrum yT. mentagrophytes. La PCR multiplex fue positiva para dermatofitos en 19 pacientes, superó el cultivo micológico por 11 muestras, detectando secuencias del T. mentagrophytes y otros dermatofitos,que fueron negativos al cultivo micológico. Se obtuvo una sensibilidad del 97% y especificidad del 73,2%. Los resultados obtenidos en este trabajo demuestran que la PCR multiplex puede detectar especies que no son aisladas en cultivo, logrando mejorar el diagnóstico y la toma de decisiones para el tratamiento en las infecciones fúngicas.
Abstract
Tinea unguiumis a superficial fungal infection due to dermatophytes that affects the ungueal plate of fingers and toes. It affects more than 10% of the population throughout the world with an increasing prevalence in the last years. Conventional diagnostic methods such as direct examination and mycological and fungal culture are of low specificity and sensibility respectively. Recent advances in DNA sequencing have allowed direct identification of dermatophytes with quick results in 24-48 hours. The aim of this study was to compare the effectiveness of PCR multiplex vstraditional methods in the diagnosis of dermatophyte infection of the ungueal plate, and to determine future parameters for its standardization. Forty-nine patients with diagnosis of tinea unguium and 10 samples of healthy control individuals were evaluated. Conventional techniques and PCR multiplex were applied to all samples to identify common DNA sequences of dermatophytes and specific sequences of T. rubrumand T. mentagrophytes. PCRm was positive for dermatophytes in 19 patients, overcoming the mycological culture by 11 samples, detecting the sequence of T. mentagrophytesand other dermatophytes, which were negative to the mycological culture. A high sensitivity of 97% and a specificity of 73.2% were obtained. The results obtained in this work show that multiplex PCR can detect species that are not isolated in culture, achieving better diagnosis and decision making for treatment in fungal infections.
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