基于重组酶聚合酶扩增-CRISPR策略的电化学生物传感器用于海月水母环境DNA的快速检测OA
An Electrochemical Biosensor Based on Recombinase Polymerase Amplification-CRISPR Strategy for Rapid Detection of Aurelia spp.Environmental DNA
海月水母暴发引起的核电冷源系统堵塞会带来潜在的核安全运行风险.然而,现有的核电冷源生物监测技术在现场快速监测与早期预警方面仍存在明显短板.本研究基于重组酶聚合酶扩增(RPA)和规律间隔成簇短回文重复序列(CRISPR)相结合的策略,构建了针对海月水母环境DNA(eDNA)的高特异性电化学生物传感快速检测体系.研究结果表明,此传感器可在45 min内完成从核酸识别到电化学信号读出的全流程检测,同时具有优异的分析性能,包括较宽的检测范围(10～1010 copy/μL)、较低的检出限(1.89 copy/μL)和定量限(5.62 copy/μL).通过对比分析本传感器和荧光定量聚合酶链式反应(PCR)扩增技术对水族箱样本的测试结果,验证了本传感器检测海月水母eDNA的准确性,Bland-Altman分析结果表明,两种方法的检测结果一致.在实际样本检测中,本传感器的检测结果与生态调查结果相吻合.本研究开发的电化学生物传感器可为核电冷源领域海月水母的现场快速监测提供新的检测技术,具有良好的工程应用前景.
The outbreak of the Aureliaspp.can cause potential safety risks by blocking the cooling water intake of nuclear power plants.However,current biological monitoring methods are unable to meet the requirements of rapid monitoring and early warning.In this study,an electrochemical biosensor based on recombinase polymerase amplification(RPA)and clustered regularly interspaced short palindromic repeats(CRISPR)was developed for rapid detection of Aurelia spp.environmental DNA(eDNA).The results demonstrated that the biosensor enabled signal readout within 45 min and exhibited excellent analytical performance,including a wide detection range(10‒1010 copy/μL),low limit of detection(1.89 copy/μL)and limit of quantitation(5.62 copy/μL).The comparison of the testing results of aquarium samples provided by the biosensor and real-time quantitative PCR confirmed the accuracy of the biosensor,while Bland-Altman analysis further confirmed the agreement between the two methods.In actual sample analysis,the detection results obtained by the biosensor were consistent with those from ecological surveys.The developed biosensor demonstrated promising potential for application in monitoring Aurelia spp..
叶长睿;张朝鑫;吴可欣;张曼;王少鹏;张翠忠;王丽伟
广西大学海洋学院,广西南海珊瑚礁研究重点实验室,南宁 530004广西大学化学化工学院,南宁 530004广西大学海洋学院,广西南海珊瑚礁研究重点实验室,南宁 530004广西大学海洋学院,广西南海珊瑚礁研究重点实验室,南宁 530004广西大学海洋学院,广西南海珊瑚礁研究重点实验室,南宁 530004广西民族师范学院化学与材料学院,崇左 532200广西大学海洋学院,广西南海珊瑚礁研究重点实验室,南宁 530004
海月水母环境DNA重组酶聚合酶扩增规律间隔成簇短回文重复序列电化学生物传感器
Aurelia spp.Environmental DNARecombinase polymerase amplificationClustered regularly interspaced short palindromic repeatsElectrochemical biosensor
《分析化学》 2026 (8)
1389-1399,中插28-中插30,14
国家自然科学基金项目(No.22564003)、广西八桂青年拔尖人才项目(No.GXR-2BGQ2525028)和广西民族师范学院高层次人才项目(No.2025XBNGCC27)资助. Supported by the National Natural Science Foundation of China(No.22564003),the Guangxi Bagui Talent Project(No.GXR-2BGQ2525028)and the High-Level Talent Project of Guangxi Minzu Normal University(No.2025XBNGCC27).
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