title: Comparative detection of a large population of grapevine viruses by TaqMan®RT-qPCR and ELISA.
abstract: Grapevine (Vitis spp.) can be infected by numerous viruses that are often widespread and of great economic importance. Reliable detection methods are necessary for sanitary selection which is the only way to partly control grapevine virus diseases. Biological indexing and ELISA are currently the standard methods for screening propagation material, and PCR-methods are becoming increasingly popular. Due to the diversity of virus isolates, it is essential to verify that the tests allow the detection of the largest possible virus populations. We developed three quadruplex TaqMan®RT-qPCR assays for detecting nine different viruses that cause considerable damage in many vineyards world-wide. Each assay is designed to detect three viruses and the grapevine Actin as an internal control. A large population of grapevines from diverse cultivars and geographic location was tested for the presence of nine viruses: Arabis mosaic virus (ArMV), Grapevine fleck virus (GFkV), Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated viruses (GLRaV-1, -2, -3), Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine virus A (GVA), and Grapevine virus B (GVB). In general, identical results were obtained with multiplex TaqMan®RT-qPCR and ELISA although, in some cases, viruses could be detected by only one of the two techniques.
authors.affiliation: Université de Haute-Alsace, Laboratoire Vigne Biotechnologies & Environnement, 33 rue de Herrlisheim, 68100, Colmar, France; SEDIAG SAS Company, Technopôle Agro-Environnement, RD 31, 21110 Bretenière, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Bruisson
authors.affiliation: Université de Haute-Alsace, Laboratoire Vigne Biotechnologies & Environnement, 33 rue de Herrlisheim, 68100, Colmar, France; SEDIAG SAS Company, Technopôle Agro-Environnement, RD 31, 21110 Bretenière, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Lebel
authors.affiliation: Université de Haute-Alsace, Laboratoire Vigne Biotechnologies & Environnement, 33 rue de Herrlisheim, 68100, Colmar, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Walter
authors.affiliation: SEDIAG SAS Company, Technopôle Agro-Environnement, RD 31, 21110 Bretenière, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Prevotat
authors.affiliation: SEDIAG SAS Company, Technopôle Agro-Environnement, RD 31, 21110 Bretenière, France.
authors.forename: Sam
authors.initials: S
authors.lastname: Seddas
authors.affiliation: Université de Haute-Alsace, Laboratoire Vigne Biotechnologies & Environnement, 33 rue de Herrlisheim, 68100, Colmar, France. Electronic address: paul.schellenbaum@uha.fr.
authors.forename: Paul
authors.initials: P
authors.lastname: Schellenbaum
mesh_terms: D029381:Closteroviridae / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D017931:DNA Primers; D018076:DNA, Complementary; D004797:Enzyme-Linked Immunosorbent Assay; D052677:Flexiviridae / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D017860:Nepovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D010935:Plant Diseases / Q000821:virology; D012367:RNA, Viral / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D046830:Tymoviridae / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D027843:Vitis / Q000821:virology*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral
keywords: Molecular detection; Multiplex; Serological detection; Vitis