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首页> 外文会议>Frontiers in biological detection: from nanosensors to systems VI >Detection of Target DNA using Photo-Reactive Protoporphyrin Moeity on a Nanocomposite Substrate
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Detection of Target DNA using Photo-Reactive Protoporphyrin Moeity on a Nanocomposite Substrate

机译：在纳米复合基质上使用光反应原卟啉部分检测靶DNA

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Detection of pathogens from infected biological samples through conventional process involves cell lysis and purification. The main objective of this work is to minimize the time and sample loss, as well as to increase the efficiency of detection of biomolecules. Electrical lysis of medical sample is performed in a closed microfluidic channel in a single integrated platform where the downstream analysis of the sample is possible. The device functions involve, in a sequence, flow of lysate from lysis chamber passed through a thermal denaturation counter where dsDNA is denatured to ssDNA, which is controlled by heater unit. A functionalized binding chamber of ssDNA is prepared by using ZnO nanorods as the matrix and functionalized with bifunctional carboxylic acid, 16-(2-pyridyldithiol) hexadecanoic acid (PDHA) which is further attached to a linker molecule 1-ethyl-3-(3-dimethylaminopropyl) (EDC). Linker moeity is then covalently bound to photoreactive protoporphyrin (PPP) molecule. The photolabile molecule protoporphyrin interacts with -NH_2 labeled single stranded DNA (ssDNA) which thus acts as a probe to detect complimentary ssDNA from target organisms. Thereafter the bound DNA with protoporphyrin is exposed to an LED of particular wavelength for a definite period of time and DNA was eluted and analyzed. UV/Vis spectroscopic analysis at 260/280 nm wavelength confirms the purity and peak at 260 nm is reconfirmed for the elution of target DNA. Quantitative and qualitative data obtained from the current experiments show highly selective detection of biomolecule such as DNA which have large number of future applications in Point-of-Care devices.
机译：通过常规方法从感染的生物样品中检测病原体涉及细胞裂解和纯化。这项工作的主要目的是最大程度地减少时间和样品损失，并提高生物分子的检测效率。在单个集成平台中，可以在封闭的微流体通道中进行医学样品的电裂解，在该平台中可以进行样品的下游分析。该设备的功能依次涉及从裂解室通过热变性计数器的裂解液流，在该计数器中，dsDNA变性为ssDNA，由加热器单元控制。通过使用ZnO纳米棒作为基质并用双官能羧酸16-（2-吡啶基二硫醇）十六烷酸（PDHA）官能化来制备ssDNA的功能化结合室，该双官能团羧酸进一步连接至连接分子1-乙基-3-（3 -二甲基氨基丙基）（EDC）。然后，连接基部分与光反应性原卟啉（PPP）分子共价结合。光不稳定分子原卟啉与-NH_2标记的单链DNA（ssDNA）相互作用，因此可作为探针来检测来自靶标生物的互补ssDNA。然后，将与原卟啉结合的DNA暴露于特定波长的LED一定时间，然后洗脱并分析DNA。在260/280 nm波长处的UV / Vis光谱分析证实了纯度，并再次确认了在260 nm处的峰可洗脱目标DNA。从当前实验中获得的定量和定性数据显示出对生物分子（如DNA）的高度选择性检测，这些分子在护理点设备中具有大量的未来应用。

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