Molecular identification tools (LAMP)

Description

LAMP (Loop-mediated isothermal Amplification) is a simple technique for identification based on the amplification of DNA at a constant temperature using pre-selected sets of primers. The primers are considered unique for a particular taxon or taxon group. The amplification product is then detected through photometry, usually by a fluorescence technique.

Within the framework of the project FF-IPM, different sets of LAMP primers are developed, targeting different representatives of two genera: Ceratitis and Bactrocera.

For Ceratitis, in addition to the target species Ceratitis capitata, the emphasis is of related species that are also considered of economic significance (as agricultural pests): Ceratitis FARQ complex (i.e.fasciventris, anonae, rosa, quilicii), and Ceratitis cosyra. To effectively
differentiate between these Ceratitis species, based on mitochondrial cox1 and cob genes, specific Ceratitis LAMP primer sets were designed and screened. 986 homologous sequences of 58 species of Ceratitis from GenBank and BOLDSYSTEM were served as negative controls to guarantee the high specificity of the primers. The experimental conditions optimization results showed that F3/B3:FIP/BIP=1:8 was the optimal primer concentration ratio, and 63°C was the optimal reaction temperature. A similar approach is under development for representatives of the genus Bactrocera, where the focus is on differentiating, primarily,between B. dorsalis, B. zonata, B. latifrons and B. oleae.