title: Simultaneous detection of three lily-infecting viruses using a multiplex Luminex bead array.
abstract: A Luminex bead array was applied to detect multiple-virus coinfection in lily plants exhibiting typical symptoms, and the efficiency of this detection system was assessed. Specific primer sets for the simultaneous detection of 4 targets in virus-infected lily plants were constructed and used for reverse transcription (RT)-polymerase chain reaction (PCR), and specific probes were used for Luminex-based assay. Each of the 4 targets was amplified, and the amplicons were used for Luminex bead array experiments. A Luminex bead array analysis of lily-infecting viruses was performed using the quadruplex RT-PCR products followed by hybridization between the biotinylated targets and anti-tagged microsphere beads. The hybridization products produced fluorescence signals that were detected by the Luminex system. Signal strengths were analyzed by their median fluorescence intensity (MFI) values. Detection of the different target elements was found to be very specific to the corresponding viruses in lilies, and coinfection with multiple viruses was specifically detected via the MFI signals. Therefore, the use of a Luminex bead array for the detection of co-infected multiple viruses in lily plants can be an improved system for screening and analyzing multiple-virus infection.
authors.affiliation: Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, Seoul 139-774, Republic of Korea.
authors.forename: Mi Sang
authors.initials: MS
authors.lastname: Lim
authors.affiliation: Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, Seoul 139-774, Republic of Korea.
authors.forename: Su Min
authors.initials: SM
authors.lastname: Kim
authors.affiliation: Plant Virus Genbank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University, Seoul 139-774, Republic of Korea. Electronic address: paxs@swu.ac.kr.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Choi
mesh_terms: D060085:Coinfection / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D005453:Fluorescence; D027762:Lilium / Q000821:virology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D010942:Plant Viruses / Q000302:isolation & purification*; D012328:RNA Viruses / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers
keywords: Coinfection; Lily-infecting virus; Luminex bead array; Multiplex detection