Probe-based quantitative PCR and RPA-Cas12a molecular diagnostics for detection of the tomato pest Phthorimaea absoluta (Lepidoptera: Gelechiidae)

Kyle M. Lewald; Wenqi Song; Daniel Eweis-LaBolle; Cindy Truong; Kristine E. Godfrey; Joanna C. Chiu

doi:10.1093/jee/toad079

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Kyle M. Lewald,¹ Wenqi Song,¹ Daniel Eweis-LaBolle,¹ Cindy Truong,¹ Kristine E. Godfrey,² Joanna C. Chiu^1,*

¹Department of Entomology and Nematology, College of Agricultural and Environmental Sciences, University of California Davis, One Shields Ave, Davis, CA 95616, USA
²Contained Research Facility, University of California, Davis, 555 Hopkins Rd, Davis, CA 95616, USA

^*Corresponding author, mail: jcchiu@ucdavis.edu.

Abstract

The tomato pest Phthorimaea absoluta Meyrick is highly invasive but has not yet invaded North America. However, several morphologically similar species are already present, making detection of P. absoluta presence and invasion challenging. We designed a quantitative PCR molecular diagnostic to differentiate P. absoluta, P. operculella (Zeller), or Keiferia lycopersicella (Walsingham) (Lepidoptera: Gelechiidae) DNA. Additionally, we developed an RPA-Cas12a molecular diagnostic that allows for the isothermal detection of P. absoluta DNA, eliminating the need for a thermocycler. The results of the RPA-Cas12a diagnostic can be visualized simply using a UV light source and cell phone camera. We expect these diagnostics to improve quarantine and prevention measures against this serious agricultural threat.

Citation Download Citation

Kyle M. Lewald, Wenqi Song, Daniel Eweis-LaBolle, Cindy Truong, Kristine E. Godfrey, and Joanna C. Chiu "Probe-based quantitative PCR and RPA-Cas12a molecular diagnostics for detection of the tomato pest Phthorimaea absoluta (Lepidoptera: Gelechiidae)," Journal of Economic Entomology 116(3), 993-1001, (13 May 2023). https://doi.org/10.1093/jee/toad079

Received: 28 December 2022; Accepted: 22 April 2023; Published: 13 May 2023