The invasive Mediterranean fanworm Sabella spallanzanii (Gmelin, 1791) is a notifiable organism under New Zealand’s Biosecurity Act and is recognized as a marine pest of particular concern, that must be reported to the Ministry for Primary Industries (MPI), New Zealand. Since its first detection in 2008, great effort and financial resources are put into surveillance and removal of individuals to contain population growth and spread. Sensitive molecular detection techniques gain great interest and are being increasingly tested for the fanworm detection in marine high-risk sites (i.e., ports and marinas) around New Zealand. However, conventional molecular detection via PCR assays from environmental DNA (eDNA) samples requires specific laboratory resources and technical expertise. This restricts the wider applicability of this approach by biosecurity practitioners or communities willing to be engaged in biosecurity surveillance. To provide end-users with a fast, easy and highly specific way to detect S. spallanzanii directly at the site of interest, a species-specific recombinase polymerase amplification (RPA) assay was designed to be read-out with lateral flow strips (RPA-LF). The RPA generates amplification within 20 minutes at 37-39°C, with a detection limit of 10 pg of the target DNA and was matching the detection limit of digital droplet PCR (ddPCR) when performed on eDNA samples. A simplified visual protocol for non-scientist users of the assay was developed and improved through independent trials with different end-user groups. The assay applicability was verified in a final validation trial with participants without scientific background resulting in 50 percent of the participants successfully detecting S. spallanzanii. Participants rated the ease of use and performance and read-out mostly as easy-to-very easy with overall positive written feedback on its usability for citizen science applications.

侵入性地中海扇虫鲭鱼(Gmelin, 1791) 是新西兰《生物安全法》规定的应通报生物，被认为是一种特别关注的海洋有害生物，必须向新西兰初级产业部 (MPI) 报告。自 2008 年首次发现以来，我们投入了大量精力和财力来监视和清除个人，以遏制人口增长和传播。敏感的分子检测技术引起了极大的兴趣，并且越来越多地在新西兰周围的海洋高风险地点（即港口和码头）进行扇虫检测的测试。然而，传统的分子检测通过环境 DNA (eDNA) 样本的 PCR 检测需要特定的实验室资源和技术专长。这限制了生物安全从业者或愿意参与生物安全监测的社区更广泛地适用这种方法。为最终用户提供一种快速、简单且高度特异性的检测方式S. spallanzanii直接在感兴趣的位点，物种特异性重组酶聚合酶扩增 (RPA) 测定被设计为用横向流动条 (RPA-LF) 读出。RPA 在 37-39°C 下在 20 分钟内产生扩增，目标 DNA 的检测限为 10 pg，并且在对 eDNA 样本进行时与数字微滴 PCR (ddPCR) 的检测限相匹配。通过与不同最终用户组的独立试验，开发并改进了针对该测定的非科学家用户的简化视觉协议。该检测方法的适用性在最终验证试验中得到验证，参与者没有科学背景，导致 50% 的参与者成功检测S. spallanzanii. 参与者对易用性、性能和读取的评价大多为易于非常容易，并对其在公民科学应用中的可用性进行了总体正面的书面反馈。