title: Evaluation of a fluorogenic real-time reverse transcription-polymerase chain reaction method for the specific detection of all known serotypes of porcine teschoviruses.
abstract: Performance of a real-time reverse-transcription polymerase chain reaction method for the rapid, simple and reliable detection of porcine teschovirus (PTV) was assessed. The method was based on the use of a set of oligonucleotides consisting of two specific primers and a fluorogenic TaqMan-MGB probe. Reverse transcription and PCR reactions were performed sequentially in one step. As a result the whole procedure was simple and rapid, taking less than 3h for completion. The method reacted in a dose-dependent manner with prototype strains for the eleven known PTV serotypes (PTV1-11), with higher analytical sensitivity than other gel-based RT-PCR methods described, which were performed in parallel to allow for a comparison. The assay did not cross-react with other related viruses or porcine viruses tested. The diagnostic performance of the method was analyzed using a panel of field samples consisting of pig fecal and pig slurry samples. As a conclusion, this technique is adequate and convenient for porcine teschovirus detection, both for diagnosis as well as in environmental investigations.
authors.affiliation: Centro de Investigación en Sanidad Animal-INIA, Ctra. Algete-El Casar s/n, 28130 Valdeolmos, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Cano-Gómez
authors.forename: Dolores
authors.initials: D
authors.lastname: Buitrago
authors.forename: Jovita
authors.initials: J
authors.lastname: Fernández-Pinero
authors.forename: Paloma
authors.initials: P
authors.lastname: Fernández-Pacheco
authors.forename: Carmen
authors.initials: C
authors.lastname: Mansilla
authors.forename: Montserrat
authors.initials: M
authors.lastname: Agüero
authors.forename: Miguel Angel
authors.initials: MA
authors.lastname: Jiménez-Clavero
mesh_terms: D000818:Animals; D017931:DNA Primers; D010850:Picornaviridae Infections / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping; D013552:Swine; D013553:Swine Diseases / Q000453:epidemiology / Q000821:virology*; D052700:Teschovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D014871:Water Microbiology; D014876:Water Pollution
chemical_list: D017931:DNA Primers; D012367:RNA, Viral