eDNA技术用CRISPR精确追踪致命水母

IF 7.3 2区 环境科学与生态学 Q1 ECOLOGY Ecological Informatics Pub Date : 2025-05-01 Epub Date: 2025-01-15 DOI:10.1016/j.ecoinf.2025.103008
Maslin Osathanunkul
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引用次数: 0

摘要

沿海地区的公共安全有赖于对危险箱形水母,特别是布氏箱形水母的检测和监测。CRISPR-Cas12a技术用于创建和验证一种新的基于eDNA的检测技术,用于快速、敏感和准确地鉴定泰国湾的C. buitendijki eDNA。在分析了63个位点的567个反应后,CRISPR-Cas12a成功地在17个位点检测到eDNA,而早期的方法没有成功。有趣的是,单个水样和每个位点的一个CRISPR-Cas12a复制可以达到95%或更高的检出率,证明了该方法的有效性。该方法的准确性在现场样品采集和实验室分析阶段都很明显,因为它具有低假阳性率和高真阳性率。尽管CRISPR-Cas12a eDNA方法不需要昂贵的科学仪器,但它不能定量样品中的eDNA,只能简单地报告其存在或不存在。尽管存在这种限制,该方法达到了每个反应0.15个拷贝的检测限(LOD),通过硅和体外测试验证了其强大的特异性。CRISPR-Cas12a的有效性、成本效益和易使用性使其对发展中国家特别有利。该方法为早期预警和预防措施提供了有力的工具,加强了海滩安全协议,并减轻了与水母接触有关的风险。它的可扩展性和可负担性支持常规监测,促进可持续沿海管理,并保护依赖旅游业的经济。CRISPR-Cas12a eDNA系统通过将科学严谨性与实用性结合起来,加强了海洋生物多样性监测,为公共卫生和环境管理的进步铺平了道路。
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eDNA tech tracks lethal jellyfish with CRISPR precision
Coastal public safety depends on detecting and monitoring dangerous box jellyfish, especially Chiropsoides buitendijki. CRISPR-Cas12a technology was used to create and validate a new eDNA-based detection technique for rapid, sensitive, and accurate identification of C. buitendijki eDNA in the Gulf of Thailand. After analysing 567 reactions across 63 sites, CRISPR-Cas12a successfully detected eDNA at 17 locations where earlier methods had been unsuccessful. Interestingly, a single water sample and one CRISPR-Cas12a replicate per site could achieve 95 % or higher detection rates, proving the method's efficacy. This method's accuracy was evident in both field sample collection and laboratory analysis stages, as it exhibited a low false positive rate and a high true positive rate. Although the CRISPR-Cas12a eDNA method does not require expensive scientific instruments, it cannot quantify eDNA in samples, simply reporting its presence or absence. Despite this limitation, the method achieved a detection limit (LOD) of 0.15 copies per reaction, with robust specificity validated through both in silico and in vitro testing. CRISPR-Cas12a's effectiveness, cost-efficiency, and ease to use make it particularly advantageous for developing countries. The method offers a powerful tool for early warning and preventive measures, enhancing beach safety protocols, and mitigating risks associated with jellyfish encounters. Its scalability and affordability support routine monitoring, promoting sustainable coastal management, and protecting tourism-dependent economies. The CRISPR-Cas12a eDNA system enhances marine biodiversity monitoring by integrating scientific rigour with practical utility, paving the way for advancements in public health and environmental management.
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来源期刊
Ecological Informatics
Ecological Informatics 环境科学-生态学
CiteScore
8.30
自引率
11.80%
发文量
346
审稿时长
46 days
期刊介绍: The journal Ecological Informatics is devoted to the publication of high quality, peer-reviewed articles on all aspects of computational ecology, data science and biogeography. The scope of the journal takes into account the data-intensive nature of ecology, the growing capacity of information technology to access, harness and leverage complex data as well as the critical need for informing sustainable management in view of global environmental and climate change. The nature of the journal is interdisciplinary at the crossover between ecology and informatics. It focuses on novel concepts and techniques for image- and genome-based monitoring and interpretation, sensor- and multimedia-based data acquisition, internet-based data archiving and sharing, data assimilation, modelling and prediction of ecological data.
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