Sensitive environmental DNA detection via lateral flow assay (dipstick) - A case study on corallivorous crown-of-thorns sea star (Acanthaster cf. solaris) detection

Detection of low-density CoTS populations is critical for early warning of outbreaks and can lead to early management interventions (e.g., culling). A lateral flow assay method for eDNA detection was developed using a commercially available nucleic acid lateral flow device (PCRD™) in combination with a previously developed species-specific mtDNA primer. This method was applied to eDNA water samples collected in field locations where CoTS were present in low (=nonoutbreak) population densities to demonstrate the sensitivity of the method to detect these populations.Aquarium raised 6- to 12-month-old juvenile CoTS were used for laboratory testing to prepare dilution series of genomic DNA.A total of 58 water samples were collected from two locations on the Great Barrier Reef (GBR), Australia; 40 samples from Big Vicky's Reef in August 2019 and 18 samples from Elizabeth Reef in May 2019. See Doyle and Uthicke (2021) for full sample processing procedures.Each field sample was analysed via ddPCR and PCR/LFA and assay sensitivity was compared as well as an assessment of the cost and time to complete sample analysis.All data associated with this study can be found either in the main text or appendices of Doyle and Uthicke (2021) linked below.

低密度棘冠海星（Crown-of-Thorns Starfish, CoTS）种群的检测，对于其暴发的早期预警至关重要，同时可为早期管理干预（如灭杀作业）提供决策依据。
本研究借助市售核酸侧向流检测设备（PCRD™），结合此前开发的物种特异性线粒体DNA（mitochondrial DNA, mtDNA）引物，构建了一种用于环境DNA（environmental DNA, eDNA）检测的侧向流分析方法。将该方法应用于野外采集的水环境DNA样本，这些样本取自低密度（即非暴发阶段）CoTS种群存在的区域，以验证该方法对这类种群的检测灵敏度。
本研究使用水族箱饲养的6至12月龄幼年CoTS开展实验室测试，以制备基因组DNA的梯度稀释系列。
共计58个水样采集自澳大利亚大堡礁（Great Barrier Reef, GBR）的两个区域：2019年8月采集自大维姬礁的40个样本，以及2019年5月采集自伊丽莎白礁的18个样本。完整的样本处理流程详见Doyle与Uthicke（2021）的研究成果。
所有野外样本均通过数字液滴PCR（droplet digital PCR, ddPCR）与PCR/侧向流分析（PCR/LFA）完成检测，同时比较了两种检测方法的灵敏度，并评估了完成样本分析所需的成本与耗时。
本研究相关的全部数据，均可在下文链接的Doyle与Uthicke（2021）论文的正文或附录中获取。