Multiplex loop‐mediated isothermal amplification assay for the identification of three major whitefly species in the greenhouse

A multiplex loop‐mediated isothermal amplification (mLAMP) assay was developed for the identification of three species of whitefly, Trialeurodes vaporariorum, Bemisia tabaci Middle East‐Asia Minor 1 (MEAM1) and Mediterranean (MED), major pests in the greenhouse. Each of the specific LAMP primer sets was designed based on the mitochondrial cytochrome oxidase I (mtCOI) gene sequence. The mLAMP reactions using primer mixtures labelled with fluorescent dye were performed at 63°C for 60 min and centrifuged with polyethyleneimine. Thus, T. vaporariorum, MEAM1 and MED were clearly identified by the colour precipitates under UV light. The mLAMP procedure described in this study is cost‐effective and can be performed in the field not only in the laboratory, because this method is a single analysis and does not need a special gene amplification device.