Multi-Target Strategy for Pan/Foot-and-Mouth Disease Virus (FMDV) Detection : A Combination of Sequences Analysis, in Silico Predictions and Laboratory Diagnostic Evaluation
Rios, Liliam (University of New Brunswick. Reiman Cancer Research Laboratory)
Perera, Carmen L. (Centro Nacional de Sanidad Agropecuaria (La Habana, Cuba))
Coronado, Liani (Centro Nacional de Sanidad Agropecuaria (La Habana, Cuba))
Relova, Damarys (Centro Nacional de Sanidad Agropecuaria (La Habana, Cuba))
Álvarez, Ana M. (Instituto Nacional de Investigaciones Agricolas)
Ganges, Llilianne (Institut de Recerca i Tecnologia Agroalimentàries. Centre de Recerca en Sanitat Animal)
Díaz de Arce, Heidy (Hospital Italiano de Buenos Aires (Argentina))
Núñez, J. Ignacio (Institut de Recerca i Tecnologia Agroalimentàries. Centre de Recerca en Sanitat Animal)
Pérez, Lester Josue (Dalhousie University. Dalhousie Medicine New Brunswick)

Fecha:	2018
Resumen:	Foot-and-mouth disease (FMD) is a highly contagious viral disease affecting cloven-hoofed animals that causes severe economic losses. The disease is characterized by a vesicular condition and it cannot be differentiated from other vesicular diseases. Therefore, laboratory confirmation of any suspected FMD case is compulsory. Despite viral isolation in cell cultures has been considered for many years as the gold standard for FMD diagnosis, the advantages of real-time reverse transcription polymerase chain reaction (rRT-PCR) technology have motivated its use directly in clinical specimens for FMD diagnosis. The current work was aimed to develop and validate a molecular multi-check strategy using rRT-PCR (mMulti-rRT-PCR) based on SYBR-Green I for pan/foot-and-mouth disease virus (pan/FMDV) diagnosis. From in silico approaches, different primer pairs previously reported were selected and modified to reduce the likelihood of viral escape as well as potential failures in the pan/FMDV detection. The analytical parameters were evaluated using a high number of representative viral strains. The repeatability of the assay and its performance on field samples were also assessed. The mMulti-rRT-PCR was able to detect emergent FMDV strains that circulated in South America between the years 2006-2010 and on which the single rRT-PCRs failed when they were applied independently. The results obtained here showed that the proposed system is an accurate and rapid diagnosis method for sensitive and specific detection of FMDV. Thus, a validated mMulti-rRT-PCR assay based on SYBR-Green I detection coupled to melting curves resolution for pan/FMDV diagnosis on clinical samples is proposed. This study also highlights the need to incorporate the multi-target detection principle in the diagnosis of highly variable agents, specially, of those listed by OIE like FMDV.
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Foot-and-mouth disease virus ; Multi-target strategy ; Sequences analysis ; Primer efficiency evaluation ; Entropy ; Amplicon stability
Publicado en:	Frontiers in veterinary science, Vol. 5 (july 2018) , ISSN 2297-1769

DOI: 10.3389/fvets.2018.00160
PMID: 30050913

13 p, 2.8 MB

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Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias de la salud y biociencias > Centre de Recerca en Sanitat Animal (CReSA-IRTA)
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