Application of the LAMP Assay as a Diagnostic Technique for Rapid Identification of Thrips tabaci (Thysanoptera: Thripidae)

Abstract

Rapid and accurate identification of potentially invasive taxa that may cause high economic losses or environmental damage is of critical importance. The onion thrips, Thrips tabaci Lindeman, ranks as one of the world's most destructive agricultural pests and commonly found in imported agricultural products and field samples, but is prone to undetected transport because of its minute size as well as cryptic behavior. Although traditional taxonomic methods are pretty useful in straightforward assignment of specimens to the genus Thrips, identification in the species level is much more difficult and requires expertise, knowledge, and experience. Furthermore, it is often difficult or impossible to identify or distinguish this species from other thrips by using material from other stages of development. Based on the foregoings, use of a molecular technique known as loop-mediated isothermal amplification (LAMP) as a rapid and robust alternative species diagnostic tool would be valuable. In this study, a relatively quick and simple method was used to detect the presence of onion thrips DNA rapidly and discriminate it from other species, by using material from different stages of development. Not only LAMP itself required less than 1 h to complete but also amounts of DNA as little as that recovered from a single specimen were adequate for the detection. Another advantage of this identification system is that nonspecialists will be able to make faster and cheaper identifications.

Simple and on‐site DNA purification for LAMP reaction applicable to non‐adult tephritid fruit fly (Diptera: Tephiritidae)

“…Although this method has often been used to detect the disease phytophthoras and viruses that infect plants (Fukuta et al., 2005; Suzuki & Miyake, 2017), no report describes its application to insects. This sampling procedure presents benefits of lower labour and time costs than the conventionally used procedure, which uses crushed insect bodies in buffer (Fekrat et al, 2015) or distilled water (Przybylska, Fiedler, Kucharczyk, & Obrępalska‐Stęplowska, 2015). For this study, a sterilized toothpick was used to pierce a living larva.…”

Development of a loop-mediated isothermal amplification (LAMP) assay for the identification of the invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) from frass

“…Among the most versatile, sensitive and specific methods, loop-mediated isothermal amplification (LAMP) can be used as a field-friendly and cost-effective diagnostic tool (Notomi et al 2000 , 2015 ). Several LAMP tests have been used both in the field and in laboratories, in particular for human and animal diseases (Lucchi et al 2010 ), in food safety controls (Abdulmawjood et al 2014 ), as well as in identifying plant pathogens (Aglietti et al 2019 ; Luchi et al 2020 ; Blaser et al 2018 ) and invasive insect pests (Huang et al 2009 ; Hsieh et al 2012 ; Fekrat et al 2015 ; Przybylska et al 2015 ; Ide et al 2016a , b ; Blaser et al 2018 ; Sabahi et al 2018 ; Rizzo et al 2020b ). LAMP is a highly specific and robust identification method for species with previously known DNA or RNA sequences and suitable for on-site application because it can be performed in a laboratory-free environment after minimal training (Kogovšek et al 2015 ).…”

Loop‐mediated isothermal amplification (LAMP) for the identification of invasive Aedes mosquito species

“…The reason for this non-specificity in the assays using hydroxy naphtol blue as a colourimetric indicator and why this can be remediated by heat treatment is not clear. Two other LAMP assays for the identification of insects [Thrips tabaci (Lindeman) (Thysanoptera: Thripidae)], cucumber fruit fly Dacus ciliatus Loew (Diptera: Tephritidae)] used crude homogenates (from thoraces or whole insects) without encountering this effect (Fekrat et al, 2015;Sabahi et al, 2018). In these studies, calcein was used as a dye indicator, which turns from orange to yellow-green after successful amplification, although this colour change can be difficult to assess and confirmation of the results by additional analyses by gel electrophoresis (as was conducted in these previous studies) is not suitable for a simple field assay.…”

“…Over the last decade, numerous LAMP assays have been developed for genetic analyses, not only mainly for the diagnosis of pathogens of medical, veterinary or agricultural importance (Bühlmann et al, 2013;Mansour et al, 2015;Li et al, 2017), but also for identifying food constituents (Focke et al, 2013) and other applications. With regard to insects, few LAMP assays have been reported for the identification of pest species (Huang et al, 2009;Hsieh et al, 2012;Fekrat et al, 2015;Przybylska et al, 2015;Ide et al, 2016;Blaser et al, 2018;Sabahi et al, 2018). The LAMP-based identification of mosquitoes has so far been developed only for the differentiation of Anopheles gambiae s.s. Giles, 1902and Anopholes arabiensis Patton, 1905(Bonizzoni et al, 2009.…”