title: [Validation of a real time RT-PCR assay to detect foot-and-mouth disease virus and assessment of its performance in acute infection].
abstract: A specific real time reverse transcription polymerase chain reaction (RT-PCRrt) for the detection of foot-and-mouth disease virus was validated using the LightCycler thermocycler 2.0 and its reagents as recommended by the World Organization for Animal Health and was assessed for the detection of the virus in acute infection of cattle experimentally vaccinated and challenged with virus A Argentina/2001 or A24 Cruzeiro. The technique proved to be robust, showing coefficients of variation lower than 4% for different ARN extractions, days or repetitions and was able to detect up to 0,4 TCID 50%, and/or up to 100 RNA molecules. In probang samples, diagnostic sensitivity was 93.1 (95% CI 86.5-96.6) and diagnostic specificity 100 (95% CI 96.3-100). The results of the challenge in vaccinated or multivaccinated bovines showed that although there were high levels of clinical protection in the vaccinated group, FMDV could be detected in all challenged groups. However, detection was 100 times lower in immunized animals.
authors.affiliation: Instituto de Virología, CICVyA-INTA Castelar, Hurlingham, Argentina. Electronic address: iviro@cnia.inta.gov.ar.
authors.forename: Norberto
authors.initials: N
authors.lastname: Fondevila
authors.affiliation: Instituto de Virología, CICVyA-INTA Castelar, Hurlingham, Argentina.
authors.forename: Diego
authors.initials: D
authors.lastname: Compaired
authors.affiliation: Dirección de Laboratorios, Servicio Nacional de Sanidad y Calidad Agroalimentaria (SENASA), Martínez, Buenos Aires, Argentina.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Maradei
authors.affiliation: Instituto de Patobiología, CICVyA-INTA Castelar, Hurlingham, Argentina.
authors.forename: Sergio
authors.initials: S
authors.lastname: Duffy
mesh_terms: D000208:Acute Disease; D000818:Animals; D001826:Body Fluids / Q000821:virology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D004947:Esophagus / Q000821:virology; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D010614:Pharynx / Q000821:virology; D012367:RNA, Viral / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D014611:Vaccination / Q000662:veterinary; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D012367:RNA, Viral; D014765:Viral Vaccines
keywords: Diagnosis; Diagnóstico; Fiebre Aftosa; Foot-and-mouth disease; RT-PCRtr; Real time RT-PCR