title: Development of a consensus reverse transcription PCR assay for the specific detection of tortoise picornaviruses.
abstract: Picornaviruses (PVs) of different terrestrial tortoise species, previously designated as Virus "X," have been frequently detected from various tissues by virus isolation in Terrapene heart cell culture as the preferred laboratory method for diagnosis. Here, we describe the development of 2 diagnostic reverse transcription (RT)-PCR-based assays for the identification and characterization of tortoise PVs belonging to the tentative genus Topivirus To test the novel diagnostic systems, PVs were isolated from swab and tissue samples collected in Germany, Italy, and Hungary between 2000 and 2013. All 25 tested isolates gave positive results with both novel consensus primer sets. Sequencing of the amplified products confirmed that all studied viruses were members of the new proposed genus Topivirus Phylogenetic analyses clearly distinguished 2 lineages within the genus. Based on sequence analysis, no association was observed between the geographic distribution and genetic relatedness. Furthermore, no strict host specificity was indicated. The PCR-based diagnosis may provide a time-saving and sensitive method to detect tortoise PVs, and evaluation of PV presence in these animals may help control virus spread.
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Marschang
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Katalin
authors.initials: K
authors.lastname: Ihász
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Renáta
authors.initials: R
authors.lastname: Kugler
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: György
authors.initials: G
authors.lastname: Lengyel
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Enikő
authors.initials: E
authors.lastname: Fehér
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Szilvia
authors.initials: S
authors.lastname: Marton
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Krisztián
authors.initials: K
authors.lastname: Bányai
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi).
authors.forename: Tara
authors.initials: T
authors.lastname: Aqrawi
authors.affiliation: LABOKLIN GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany (Marschang)Centre for Agricultural Research, Institute for Veterinary Medical Research, Hungarian Academy of Sciences, Budapest, Hungary (Ihász, Kugler, Fehér, Marton, Bányai, Farkas)Medical Centre of Hungarian Defense Forces, Force Health Laboratory Institute, Budapest, Hungary (Lengyel)FG für Umweltund Tierhygiene, University of Hohenheim, Stuttgart, Germany (Aqrawi) fszilvi@yahoo.com.
authors.forename: Szilvia L
authors.initials: SL
authors.lastname: Farkas
mesh_terms: D000818:Animals; D005060:Europe; D010802:Phylogeny; D010849:Picornaviridae / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D014426:Turtles / Q000821:virology*
chemical_list: D012367:RNA, Viral
keywords: Diagnostic reverse transcription PCR assay; Terrapene heart cell; Testudo graeca; Topivirus