摘要
核酸现场快速检测具有便捷、操作简单、无需专用实验室、报告快速等优点,能在机场、社区医院、海关、野外等各种复杂环境中进行检测。建立了一种可应用于核酸现场快速检测仪器的荧光检测系统,包括非共聚焦式的正交光路与多通道检测集成结构。前者信噪比高、小型便捷;后者切换效率高,能满足多重检测,其嵌合结构极大降低了通道间的荧光串扰。实验证明,系统的检测下限低于1.6μg/mL,荧光检测梯度性R2值(确定系数)均大于0.99,重复性测试的CV(coefficient of variance)值不超过1.27%。通道间荧光串扰测试表明,通道间无串扰问题,巨细胞病毒培养液的扩增实验准确性与稳定性较好,表明该系统能够针对多重检测进行有效荧光激发与采集。
On-site rapid detection of nucleic acid has the advantages of convenience,simple operation,no need for special laboratories,and rapid reporting,which can be detected in various complex environments such as airports,community hospitals,customs and fields.A fluorescence detection system that can be applied to onsite rapid detection instruments of nucleic acid was established,including non-confocal orthogonal optical path and multi-channel detection integrated structure.The former has the characteristics of high signal-to-noise ratio,small and convenient.The latter has high switching efficiency and can meet multiple detection,and its embedded structure greatly reduces the fluorescence crosstalk between channels.The experimental results show that the detection limit of the system is lower than 1.6μg/mL,the R2 value(coefficient of determination)of fluorescence detection gradient is greater than 0.99,and the coefficient of variance(CV)value of repeatability test does not exceed 1.27%.The inter-channel fluorescence crosstalk test shows that there is no crosstalk between channels,and the accuracy and stability of cytomegalovirus culture medium amplification experiment are good,which indicates that the system can effectively stimulate and collect fluorescence for multiple detection.
作者
杨佳羽
曾俊添
奚邦朝
刘国镪
黄绍磊
张东旭
YANG Jiayu;ZENG Juntian;XI Bangchao;LIU Guoqiang;HUANG Shaolei;ZHANG Dongxu(School of Public Health,Xiamen University,Xiamen 361102,China;State Key Laboratory of Molecular Vaccinology and Molecular Diagnostic,Xiamen University,Xiamen 361102,China;National Institute of Diagnostics and Vaccine Development in Infection Diseases,Xiamen University,Xiamen 361102,China;State Drug Administration Key Laboratory of Infectious Disease Detection Technology Research and Evaluation,Xiamen 361102,China)
出处
《应用光学》
CAS
北大核心
2023年第4期859-867,共9页
Journal of Applied Optics
基金
国家自然科学基金青年项目(62003284)
厦门大学校长基金(20720210089)。
关键词
核酸现场检测
聚合酶链式反应
荧光
实时
on-site nucleic acid detection
polymerase chain reaction
fluorescence
real-time
