title: Development of a TaqMan Probe-Based Insulated Isothermal Polymerase Chain Reaction (iiPCR) Assay for Detection of Fusarium oxysporum f. sp. cubense Race 4.
abstract: This study developed a novel and inexpensive detection method based on a TaqMan probe-based insulated isothermal polymerase chain reaction (iiPCR) method for the rapid detection of Panama disease caused by Fusarium oxysporum f. sp. cubense (Foc) race 4, which is currently among the most serious fungal vascular diseases worldwide. By using the portable POCKIT™ device with the novel primer set iiFoc-1/iiFoc-2, the Foc race 4 iiPCR assay (including DNA amplification and signal monitoring) could be completed within one hour. The developed Foc race 4 iiPCR assay is thus a user-friendly and efficient platform designed specifically for the detection of Foc race 4. The detection limit of this optimized Foc iiPCR system was estimated to be 1 copy of the target standard DNA as well as 1 fg of the Foc genomic DNA. This approach can serve as a rapid detection method for in planta detection of Foc race 4 in field-infected banana. It was concluded that this molecular detection procedure based on iiPCR has good potential for use as an efficient detection method.
authors.affiliation: Department of Plant Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Ying-Hong
authors.identifier: http://orcid.org/0000-0002-7055-0652
authors.initials: YH
authors.lastname: Lin
authors.affiliation: Department of Plant Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Yi-Jia
authors.initials: YJ
authors.lastname: Lin
authors.affiliation: Department of Plant Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Tsai-De
authors.initials: TD
authors.lastname: Chang
authors.affiliation: Department of Plant Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Li-Ling
authors.initials: LL
authors.lastname: Hong
authors.affiliation: Department of Plant Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Tzu-Yu
authors.initials: TY
authors.lastname: Chen
authors.affiliation: Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan.|Agricultural Biotechnology Center, National Chung Hsing University, Taichung, Taiwan.|NCHU-UCD Plant and Food Biotechnology Center, National Chung Hsing University, Taichung, Taiwan.
authors.forename: Pi-Fang Linda
authors.initials: PF
authors.lastname: Chang
mesh_terms: D005670:Fusarium / Q000235:genetics*; D028521:Musa / Q000382:microbiology*; D010935:Plant Diseases / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity