title: Diagnosis of Cetacean morbillivirus: A sensitive one step real time RT fast-PCR method based on SYBR(®) Green.
abstract: 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.
authors.affiliation: Departamento de Patologia - VPT, Laboratório de Patologia Comparada de Animais Selvagens - LAPCOM, Faculdade de Medicina Veterinária e Zootecnia - FMVZ, Universidade de São Paulo, Avenida Professor Orlando Marques de Paiva 87, 05508-270 São Paulo, Brazil; Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Sacristán
authors.affiliation: Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: Matilde
authors.initials: M
authors.lastname: Carballo
authors.affiliation: Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: María Jesús
authors.initials: MJ
authors.lastname: Muñoz
authors.affiliation: Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: Edwige Nina
authors.initials: EN
authors.lastname: Bellière
authors.affiliation: Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Neves
authors.affiliation: Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: Verónica
authors.initials: V
authors.lastname: Nogal
authors.affiliation: Centro de Investigación en Sanidad Animal (INIA-CISA), Ctra Algete el Casar s/n, 28130 Valdeolmos, Madrid, Spain. Electronic address: esperon@inia.es.
authors.forename: Fernando
authors.initials: F
authors.lastname: Esperón
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002591:Cetacea; D003959:Diamines; D018103:Morbillivirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D018185:Morbillivirus Infections / Q000662:veterinary* / Q000821:virology; D009930:Organic Chemicals; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014760:Viral Fusion Proteins
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D014760:Viral Fusion Proteins; C098022:SYBR Green I
keywords: Cetacean morbillivirus diagnosis; Fusion protein gene; Molecular epidemiology; Real-time RT-PCR