Development of the point-of-need detection assay for the invasive Mediterranean fanworm

The DETECT team of the Marine Biosecurity Toolbox programme is committed to equip researchers, biosecurity practitioners and prospective citizen scientists with rapid, efficient and user-friendly molecular tools for marine biosecurity surveillance. Thus, Martin Zirngibl is currently leading the development of a point-of-need assay for detecting the invasive Mediterranean fanworm Sabella spallanzanii from environmental samples. Basis of the new assay will be recombinase polymerase amplification (RPA) and the readout will be via lateral flow detection (Figure 1).Figure 1: Overview of the design and validation of the newly developed assay

Primers specific to Sabella spallanzanii were designed and tested with RPA approach. The primers that produced single strong bands on the gel were tested for specificity with three other closely related polychaetes. In order to make the newly designed assay usable in the field without requiring electricity, amplification was tested while holding the reaction tube in the hand. In the future, the assay will be transferred to read-out with a lateral flow stick.