Diagnosis of Cetacean morbillivirus A sensitive one step real time RT fast-PCR method based on SYBR® Green

Cetacean morbillivirus (CeMV) (family Paramyxoviridae, genus Morbillivirus) is considered the most pathogenic virus of cetaceans. It was first implicated in the bottlenose dolphin (Tursiops truncatus) mass stranding episode along the Northwestern Atlantic coast in the late 1980s, and in several more recent worldwide epizootics in different Odontoceti species. This study describes a new one step real-time reverse transcription fast polymerase chain reaction (real-time RT-fast PCR) method based on SYBR® Green to detect a fragment of the CeMV fusion protein gene. This primer set also works for conventional RT-PCR diagnosis. This method detected and identified all three well-characterized strains of CeMV porpoise morbillivirus (PMV), dolphin morbillivirus (DMV) and pilot whale morbillivirus (PWMV). Relative sensitivity was measured by comparing the results obtained from 10-fold dilution series of PMV and DMV positive controls and a PWMV field sample, to those obtained by the previously described conventional phosphoprotein gene based RT-PCR method. Both the conventional and real-time RT-PCR methods involving the fusion protein gene were 100- to 1000-fold more sensitive than the previously described conventional RT-PCR method. © 2015 Elsevier B.V.

Bibliographic Details

Main Authors:	Sacristán, C., Carballo Santaolalla, Matilde, Muñoz, M. J., Bellière, E. N., Neves, E., Nogal, V., Esperón Fajardo, Fernando
Format:	artículo biblioteca
Language:	English
Published:	Elsevier 2015
Subjects:	Cetacean morbillivirus diagnosis, Real-time RT-PCR, Fusion protein gene, Molecular epidemiology,
Online Access:	http://hdl.handle.net/20.500.12792/3966 http://hdl.handle.net/10261/292962
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