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Vol. 24. Núm. 8.
Páginas 500-504 (octubre 2006)
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Vol. 24. Núm. 8.
Páginas 500-504 (octubre 2006)
Originales
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PCR en tiempo real, inmunofluorescencia y cultivo para la detección de Bordetella pertussis: evaluación prospectiva y epidemiología molecular
Bordetella pertussis detection by real-time PCR, immunofluorescence and culture: prospective evaluation and molecular epidemiology
Visitas
20139
Jesús García-Martínez, Fernando Chaves
Autor para correspondencia
fchaves.hdoc@salud.madrid.org

Correspondencia: Dr. F. Chaves. Servicio de Microbiología. Hospital Universitario 12 de Octubre. Avda. Córdoba, s/n. 28041 Madrid. España.
, Efrén Salto, Joaquín R. Otero
Servicio de Microbiología. Hospital Universitario 12 de Octubre. Madrid. España
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Resumen
Bibliografía
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Estadísticas
Introducción

En los últimos años se ha observado un aumento de la incidencia de tos ferina. El objetivo de este estudio fue evaluar la aportación de varios procedimientos, incluyendo una técnica de reacción en cadena de la polimerasa (PCR) en tiempo real, al diagnóstico microbiológico de tos ferina, así como conocer la relación clonal de los aislamientos clínicos de Bordetella pertussis.

Métodos

Desde agosto de 2002 a octubre de 2003, se recogieron los exudados nasofaríngeos de pacientes pediátricos y adultos con signos clínicos de tos ferina o como parte de estudios de contactos. Estas muestras fueron procesadas para la detección de B. pertussis mediante cultivo, inmunofluorescencia directa (IFD) y PCR en tiempo real. Los aislamientos disponibles se caracterizaron molecularmente mediante electroforesis en campo pulsante (ECP).

Resultados

De las 121 muestras recogidas de 117 pacientes, se detectó B. pertussis en cultivo en 17 muestras (14,1%), con IFD en 30 (24,8%) y con PCR en 41 (33,9%). Un total de 17 aislamientos clínicos, 14 obtenidos durante el período de estudio y 3 en los años 1997, 2000 y 2001, estuvieron disponibles para ECP. Se identificaron 5 genotipos, dos de ellos (C y E) mayoritarios, con 8 y 6 aislados, respectivamente. Uno de ellos incluyó aislamientos de 1997 y 2001.

Conclusión

La PCR en tiempo real aplicada a la detección de B. pertussis proporcionó más resultados positivos que la IFD o el cultivo, pero su valor diagnóstico debe ser aclarado. Entre los aislamientos conseguidos predominaron dos clones, uno de ellos en circulación desde al menos 1997.

Palabras clave:
Bordetella pertussis
Tos ferina
PCR en tiempo real
Epidemiología molecular
Introduction

An increase in the incidence of pertussis has been observed in recent years. The aim of this study was to determine the usefulness of several procedures, including real-time PCR, for the laboratory diagnosis of pertussis, and to investigate clonal relationships among clinical isolates of Bordetella pertussis.

Methods

During the period of August 2002 to October 2003, nasopharyngeal swabs were collected from pediatric and adult patients with symptoms of pertussis, and contact cases. The samples were processed by culture, direct fluorescence assay (DFA), and real-time PCR. Most of the isolates were further characterized by pulsed-field gel electrophoresis (PFGE).

Results

Among 121 clinical samples corresponding to 117 patients, B. pertussis was detected in 17 samples by culture (14.1%), 30 samples (24.8%) by DFA and 41 samples (33.9%) by real-time PCR. Real-time PCR diagnosed 26 and 24 more cases than culture and DFA, respectively.

Seventeen clinical isolates were available for PFGE analysis, 14 collected during the study period and three in 1997, 2000 and 2001. PFGE identified 5 different genotypes, 2 of which included 8 (genotype C) and 6 (genotype E) isolates. Two of the older isolates (1997 and 2001) were identified as genotype C.

Conclusion

Real-time PCR applied to the diagnosis of pertussis provided more positive results than DFA and culture, but the true diagnostic value of these results should be clarified. Two bacterial clones were dominant, and one of them has circulated at least since 1997.

Key words:
Bordetella pertussis
Whooping cough
Real-time PCR
Molecular epidemiology
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Copyright © 2006. Elsevier España S.L.. Todos los derechos reservados
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