EPPO Bulletin (2022) 52, 141-148

Domenico Rizzo, Daniele Da Lio, Tommaso Bruscoli, Giovanni Cappellini, Linda Bartolini, Chiara Salemi, Antonio Aronadio, Dalia Del Nista, Jason B. Oliver and Elisabetta Rossi (2022)
SYBR Green-based real-time PCR test for the identification of adults and larvae of the Japanese beetle Popillia japonica Newman (Coleoptera Scarabaeidae)
EPPO Bulletin 52 (1), 141-148
Abstract: The Japanese beetle (Popillia japonica Newman) (Coleoptera Scarabaeidae) is a polyphagous invasive pest included in quarantine lists in Europe and in many countries of Africa, Asia and America. No specific molecular methods were available for the identification of adults and preimaginal stages. To bridge this gap, a rapid method (<2 h) based on SYBR Green real-time PCR for identifying larvae and adults of P. japonica was developed in this study. SYBR Green chemistry was chosen for its cost-effectiveness and adaptability although it has a lower specificity than other real-time PCR tests such as the as TaqMan test which requires a probe design. The in-silico primer specificity was tested, and the theoretical amplicon did not match any of the sequences deposited in databanks. The diagnostic specificity (exclusivity) of the test was assessed by testing the method on taxonomically related species (Popillia anomaloides Kraatz, P. hilaris Kraatz and P. fallaciosa Fairmaire), other Coleoptera and unrelated insect species, with no cross-reactions. Performance characteristics of the test, in terms of diagnostic specificity (inclusivity), analytical sensitivity, and accuracy, were evaluated. The analytical sensitivity determined on a single larva showed a LoD value of 3.20 pg/µL. The molecular test developed could be highly beneficial for the phytosanitary controls at entry points for the regulatory confirmation of specimen identity and/or for monitoring the presence and spread of P. japonica in the field.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Jason B. Oliver, Elisabetta Rossi

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
surveys/sampling/distribution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Popillia japonica		Italy