title: Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus.
abstract: A multiplex PCR method has been developed to detect, differentiate, and confirm the morphological identification of three root infecting Olpidium spp.: O. bornovanus, O. brassicae, and O. virulentus. Of the 132 root samples examined, 101 samples were infected by Olpidium spp.. Based on the morphology of resting spores, the presence of O. bornovanus was confirmed in 20.5% of the samples, whereas species identity could not be determined for the remaining samples because they failed to reproduce sexually. With multiplex PCR, it was possible to determine the Olpidium identity of all the infected samples, even when resting spores were not formed. This method was also effective for detecting Olpidium spp. in water samples. In addition, the specificity and sensitivity of multiplex PCR were evaluated. The multiplex PCR method was validated with samples of 9 different crops from 11 countries of America, Europe, and Africa.
authors.affiliation: Grupo de Virología, Instituto Agroforestal Mediterráneo, Universidad Politécnica de Valencia, Camino de Vera s/n, E-46022 Valencia, Spain. joshervs@doctor.upv.es <joshervs@doctor.upv.es>
authors.forename: José Angel
authors.initials: JA
authors.lastname: Herrera-Vásquez
authors.forename: María Del Carmen
authors.initials: Mdel C
authors.lastname: Cebrián
authors.forename: Ana
authors.initials: A
authors.lastname: Alfaro-Fernández
authors.forename: María Del Carmen
authors.initials: Mdel C
authors.lastname: Córdoba-Sellés
authors.forename: Concepción
authors.initials: C
authors.lastname: Jordá
mesh_terms: D008411:Chytridiomycota / Q000145:classification* / Q000235:genetics*; D004271:DNA, Fungal / Q000032:analysis; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D004271:DNA, Fungal