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Detection of Target DNA using Photo-Reactive Protoporphyrin Moeity on a Nanocomposite Substrate

Das, Sumana and Mishra, Madhusmita and Vasireddi, Ramakrishna and Mahapatra, Roy D (2014) Detection of Target DNA using Photo-Reactive Protoporphyrin Moeity on a Nanocomposite Substrate. In: Conference on Frontiers in Biological Detection - From Nanosensors to Systems VI, FEB 01-02, 2014, San Francisco, CA.

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Official URL: http://dx.doi.org/10.1117/12.2042556

Abstract

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 -NH2 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.

Item Type: Conference Proceedings
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Additional Information: copyright for this article belongs to SPIE-INT SOC OPTICAL ENGINEERING, 1000 20TH ST, PO BOX 10, BELLINGHAM, WA 98227-0010 USA
Keywords: ZnO nanorod; 1-ethyl-3-(3-dimethylaminopropyl) (EDC); microfluidic device; photlabile; protoporphyrin; DNA binding; automated sample preparation; cell lysis
Department/Centre: Division of Mechanical Sciences > Aerospace Engineering (Formerly, Aeronautical Engineering)
Date Deposited: 27 Jun 2014 05:39
Last Modified: 27 Jun 2014 05:39
URI: http://eprints.iisc.ernet.in/id/eprint/49388

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