Performance Evaluation of a Novel Sample In-Answer Out (SIAO) System Based on Magnetic Nanoparticles
Performance Evaluation of a Novel Sample In-Answer Out (SIAO) System Based on Magnetic Nanoparticles
复制标题
基于磁性纳米粒子的新型样本输入-应答输出 (SIAO) 系统的性能评估
DOI：
10.1166/jbn.2017.2478
复制
发表时间：
2017-12-01
影响因子：
2.9
通讯作者：
He, Nongyue
中科院分区：
文献类型：
--
作者：
Chen, Hui;Wu, Yanqi;He, Nongyue
近年来，传染病的预防、控制、诊断和治疗已成为全球公共卫生关注的焦点。传统的病原体检测方法有一些主要的缺点，包括需要高技能的工作人员和昂贵的仪器，而程序方面是复杂的，对环境敏感。这些缺点极大地限制了传统检测方法在现场病原体检测中的应用。在本文中，我们提出了一种新的基于磁性纳米颗粒的即时检测（POCT）系统，该系统可以实现病原体的样品输入-应答（SIAO）自动实时检测。对仪器进行了各种性能测试。SIAO系统与人工系统相比，核酸提取效率分别为95.49%和84.33%。在SIAO系统中通过两种方法（基于TaqMan的探针和SYBR绿色染料）的实时PCR是可实现的，具有与手动方法相当的结果。SIAO系统的核酸检测具有可重复性，优于手动检测。SIAO系统具有良好的抗污染性能，易于避免批间交叉污染。最后，使用SIAO系统进行腺病毒检测产生了与LightCycler2.0系统测定结果相似的结果。扩增图和Ct值表明，使用SIAO系统进行腺病毒检测与使用实时荧光PCR检测和商业仪器手动核酸提取后进行腺病毒检测的扩增图形状相似。总的来说，这些结果表明，使用SIAO系统进行的测试实际上等同于使用商用系统进行的手动浸提测试。
In recent years, the prevention, control, diagnosis and treatment of infectious diseases has become a global focus for public health. Traditional methods for pathogen testing have some major disadvantages including the need for highly skilled staff and expensive instrumentation, while procedural aspects are complex and sensitive to the environment. These shortcomings have greatly limited the application of traditional testing in on site pathogen detection. In this paper, we present a new point-of-care-testing (POCT) system based on magnetic nanoparticles that enable sample in-answer out (SIAO) automated real-time testing for pathogens. Various performance tests were conducted on the instrument. Nucleic acid extraction efficiencies of SIAO versus manual systems were 95.49% and 84.33%, respectively. Real-time PCR by two methods (TaqMan-based probe and SYBR green dye) in the SIAO system was achievable, with comparable results to the manual method. Nucleic acid testing with the SIAO system was repeatable and better than with manual testing. The SIAO system had good anti-pollution performance with easy avoidance of inter-assay cross contamination. Finally, use of the SIAO system for adenovirus detection produced similar results to LightCycler2.0 system assay findings. The amplification plots and Ct values suggested similar amplification plots shapes for adenovirus testing with the SIAO system and with real-time fluorescence PCR testing and commercial instrument post-manual nucleic acid extraction. Collectively, these findings indicate that testing with the SIAO system is virtually equivalent to that of manual extraction with commercial system testing.