Development of an Automated, Non-Enzymatic Nucleic Acid Amplification Test.
Development of an Automated, Non-Enzymatic Nucleic Acid Amplification Test.
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DOI：
10.3390/mi12101204
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发表时间：
2021-09-30
期刊：
影响因子：
3.4
通讯作者：
Haselton FR
中科院分区：
文献类型：
--
作者：
Zimmers ZA;Boyd AD;Stepp HE;Adams NM;Haselton FR
在核酸诊断策略中，非酶检测是最有希望应用于低资源环境护理点的方法。但是，由于灵敏度不够，这些方法仍然没有得到充分利用。受最近高灵敏度哑铃DNA扩增策略演示的启发，我们开发了一种自动化的，独立的检测目标DNA的方法。在这个新的诊断平台中，被称为自动pi驱动的环寡核苷酸转运体，磁珠捕获目标DNA，然后与其他反应溶液一起装载到微流体反应盒中。步进电机控制盒体相对于外部磁场的运动，外部磁场使磁珠自动通过反应溶液。实时荧光法用于测量哑铃在磁珠表面的积累。左撇子的DNA哑铃会产生一种独特的信号，反映非特异性扩增的水平，起到内部控制的作用。autoPiLOT检测仅检测到5fm的靶DNA，也成功地应用于S. mansoni DNA检测。autoPiLOT的设计是在开发一种敏感、用户友好、低资源、非酶诊断测试方面迈出的新一步。
Among nucleic acid diagnostic strategies, non-enzymatic tests are the most promising for application at the point of care in low-resource settings. They remain relatively under-utilized, however, due to inadequate sensitivity. Inspired by a recent demonstration of a highly-sensitive dumbbell DNA amplification strategy, we developed an automated, self-contained assay for detection of target DNA. In this new diagnostic platform, called the automated Pi-powered looping oligonucleotide transporter, magnetic beads capture the target DNA and are then loaded into a microfluidic reaction cassette along with the other reaction solutions. A stepper motor controls the motion of the cassette relative to an external magnetic field, which moves the magnetic beads through the reaction solutions automatically. Real-time fluorescence is used to measure the accumulation of dumbbells on the magnetic bead surface. Left-handed DNA dumbbells produce a distinct signal which reflects the level of non-specific amplification, acting as an internal control. The autoPiLOT assay detected as little as 5 fM target DNA, and was also successfully applied to the detection of S. mansoni DNA. The autoPiLOT design is a novel step forward in the development of a sensitive, user-friendly, low-resource, non-enzymatic diagnostic test.
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影响因子：
17.1
作者：
Xu G;Zhao H;Reboud J;Cooper JM
通讯作者：
Cooper JM
DOI：
10.1073/pnas.0407024101
发表时间：
2004-10-26
影响因子：
11.1
作者：
Dirks, RM;Pierce, NA
通讯作者：
Pierce, NA
影响因子：
1.5
作者：
HAMBURGER, J;TURETSKI, T;DERESIEWICZ, R
通讯作者：
DERESIEWICZ, R
影响因子：
6.1
作者：
Knowlton, Stephanie;Joshi, Ashwini;Tasoglu, Savas
通讯作者：
Tasoglu, Savas
影响因子：
15
作者：
Mahshid, Sahar Sadat;Camire, Sebastien;Vallee-Belisle, Alexis
通讯作者：
Vallee-Belisle, Alexis