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 degrees C for 60min 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.