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Multiplex loop‐mediated isothermal amplification assay for the identification of three major whitefly species in the greenhouse
Author(s) -
Suzuki R.,
Tanaka H.,
Suzuki K.,
Fukuta S.,
Kato S.,
Ohno T.
Publication year - 2018
Publication title -
journal of applied entomology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 60
eISSN - 1439-0418
pISSN - 0931-2048
DOI - 10.1111/jen.12522
Subject(s) - trialeurodes , loop mediated isothermal amplification , biology , greenhouse whitefly , primer (cosmetics) , multiplex , whitefly , microbiology and biotechnology , homoptera , botany , pest analysis , genetics , dna , chemistry , organic chemistry
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 ( MEAM 1) and Mediterranean ( MED ), major pests in the greenhouse. Each of the specific LAMP primer sets was designed based on the mitochondrial cytochrome oxidase I ( mt COI ) 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 , MEAM 1 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.