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A Smartphone Enabled Molecular Diagnostic Toolkit to Detect Pathogens via Isothermal Nucleic Acid Amplification on Pre-Dried Disposable Paper Strips

Masetty, Manaswini
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1627664394713446

Abstract Details

2021, MS, University of Cincinnati, Engineering and Applied Science: Chemical Engineering.
The novel Coronavirus Disease has affected the lives of millions of people all around the world, thus acquiring the need for rapid testing. Point-of-care (POC) diagnostics are such rapid tests that are used to diagnose an infectious disease, and provide the results within minutes of the test being done, thereby allowing rapid treatment. Paper-based point-of-care devices have become very popular due to their portability, low-cost and ease of fabrication and distribution. This project describes a simple and inexpensive paper-based modular system for detecting the presence/absence of DNA/RNA targets using a smartphone. Smartphones have an extremely sophisticated hardware and software package in a truly portable, user-friendly and inexpensive manner [5]. Therefore, it has played a major role in the point-of- care diagnostic field as standalone biosensing systems for medical applications [5]. They are replacing laboratory analytical and biomedical equipment. The following is a smartphone enabled nucleic acid based amplification device. Molecular based testing depends on the amplification of the target pathogen DNA/RNA. Current molecular diagnostic systems rely on Polymerase Chain Reaction (PCR) to selectively find, amplify and detect target pathogen DNA/RNA in human samples. As an alternate to PCR, Loop Mediated Isothermal Amplification (LAMP) reaction can amplify a known DNA/RNA sequence in the target pathogen without the use of bulky, power-consuming thermocyclers. Liquid aliquots can be hard to deal with and have supply chain issues, especially during a pandemic. Transportation of paper substrates is easier as all the LAMP reagents are present in the dried format on it. Paper based detection is more suitable for POC diagnostics. Paper-based LAMP platform involves a paper substrate on which the DNA/RNA amplification reaction takes place. The first aim talks about characterizing visual detection systems for detecting amplified LAMP products. This involves exploring the various methods for detecting the amplified nucleic acid target and picking the best one from the lot. The second aim proposes developing a protocol for dehydrating LAMP reagents and detecting LAMP products on paper. This entails the experimental protocol for performing amplification reaction on paper substrate. The third and final aim involves coming up with a portable platform for detection of infectious diseases. This talks about the design and working of a portable dark box which acts as a molecular diagnostic toolkit. Results and protocols are reported throughout the proposed aims to further ensure the validity of this project. Each aim has been designed after a thorough literary review of pre-existing strategies. Extensive background and insight have been provided in conjunction with a detailed experimental approach to adequately satisfy the objectives of each aim. While it is clear that LAMP on paper has immense potential for POC diagnostics, this project will build beyond that foundation to create a novel portable diagnostic toolkit for detection of pathogens. The result will be the inclusion of an isothermal heating setup in a smartphone enabled 3D printed dark box. Researchers can then build upon the findings to better optimize this toolkit for POC testing.
Aashish Priye, Ph.D. (Committee Chair)
Greg Harris, Ph.D. (Committee Member)
Surya Prasath, Ph.D. (Committee Member)
54 p.
Molecular Biology
Loop Mediated Isothermal Amplification; Smartphone Diagnostics; Point-of-Care Diagnostics; Paper Microfluidics

Recommended Citations

Citations

  • Masetty, M. (2021). A Smartphone Enabled Molecular Diagnostic Toolkit to Detect Pathogens via Isothermal Nucleic Acid Amplification on Pre-Dried Disposable Paper Strips [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1627664394713446

    APA Style (7th edition)

  • Masetty, Manaswini. A Smartphone Enabled Molecular Diagnostic Toolkit to Detect Pathogens via Isothermal Nucleic Acid Amplification on Pre-Dried Disposable Paper Strips. 2021. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1627664394713446.

    MLA Style (8th edition)

  • Masetty, Manaswini. "A Smartphone Enabled Molecular Diagnostic Toolkit to Detect Pathogens via Isothermal Nucleic Acid Amplification on Pre-Dried Disposable Paper Strips." Master's thesis, University of Cincinnati, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1627664394713446

    Chicago Manual of Style (17th edition)

ucin1627664394713446
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© 2021, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.