title: A PCR technique for the detection of Jaagsiekte sheep retrovirus in the blood suitable for the screening of ovine pulmonary adenocarcinoma in field conditions.
abstract: Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring contagious lung neoplasia caused by jaagsiekte sheep retrovirus (JSRV). Although no specific circulating antibodies against the virus can be detected in infected sheep, JSRV proviral DNA sequences can be found in peripheral blood leukocytes (PBLs) in clinically affected and in a proportion of in contact animals. In this study, existing hemi-nested PCR procedure is compared with a new one-step PCR technique that was developed to minimise potential DNA contamination and reduce sample and reagent handling. Different blood preparations were assessed and the best results were achieved on DNA prepared from buffy coat. The sensitivity of this PCR was lower in JSRV infected sheep without lesions of OPA than in clinically affected sheep, which indicate that this PCR may not be not fully appropriate for screening of individual sheep, but rather to provide results at flock level. This PCR is the only currently available blood test for detection of JSRV infected sheep and may be useful in epidemiological studies and in control programmes of OPA.
authors.affiliation: Departamento de Patología Animal, Universidad de Zaragoza, C/Miguel Server 177, 50013 Zaragoza, Spain. lasheras@unizar.es
authors.forename: Marcelo
authors.initials: M
authors.lastname: De Las Heras
authors.forename: Aurora
authors.initials: A
authors.lastname: Ortín
authors.forename: Daniela
authors.initials: D
authors.lastname: Salvatori
authors.forename: Maider
authors.initials: M
authors.lastname: Pérez de Villareal
authors.forename: Christina
authors.initials: C
authors.lastname: Cousens
authors.forename: Luis
authors.initials: L
authors.lastname: Miguel Ferrer
authors.forename: Luis
authors.initials: L
authors.lastname: Miguel Cebrián
authors.forename: José Antonio
authors.initials: JA
authors.lastname: García de Jalón
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Gonzalez
authors.forename: James
authors.initials: J
authors.lastname: Michael Sharp
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000032:analysis / Q000097:blood; D022141:Jaagsiekte sheep retrovirus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary*; D011648:Pulmonary Adenomatosis, Ovine / Q000097:blood / Q000175:diagnosis* / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis / Q000097:blood; D012756:Sheep / Q000097:blood / Q000821:virology*
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral