<p>Swift recognition of
disease-causing pathogens at the point-of-care enables life-saving treatment
and infection control. However, current rapid diagnostic devices often fail to
detect the low concentrations of pathogens present in the early stages of
infection, causing delayed and even incorrect treatments. Rapid diagnostics
that require multiple steps and/or elevated temperatures to perform have a
number of barriers to use at the point-of-care and in the field, and despite
efforts to simplify these platforms for ease of use, many still require
diagnostic-specific training for the healthcare professionals who use them.
Most nucleic acid amplification assays require hours to perform in a sterile
laboratory setting that may be still more hours from a patient’s bedside or not
at all feasible for transport in remote or low-resourced areas. The cold-chain
storage of reagents, multistep sample preparation, and costly instrumentation
required to analyze samples has prohibited many nucleic acid detection and
antibody-based assays from reaching the point-of-care. There remains a critical
need to bring rapid and accessible pathogen identification technologies that
determine disease status and ensure effective treatment out of the laboratory.</p>
<p>Paper-based diagnostics have emerged as a portable platform for antigen
and nucleic acid detection of pathogens but are often limited by their
imperfect control of reagent incubation, multiple complex steps, and
inconsistent false positive results. Here, I have developed mechanisms to
economically improve thermal incubations, automate dried reagent flow for
multistep assays, and specifically detect pathogenic antigens while improving
final output sensitivity on paper-based devices. First, I characterize
miniaturized inkjet printed joule-heaters (microheaters) that enable thermal
control for pathogen lysis and nucleic acid amplification incubation on a
low-cost paper-based device. Next, I explore 2-Dimensional Paper Networks as a
means to automate multistep visual enhancement reactions with dried reagents to
increase the sensitivity and readability of nucleic acid detection with
paper-based devices. Lastly, I aim to create a novel Reverse-Transcription
Recombinase Polymerase Reaction mechanism to amplify and detect a specific
region of the Spike protein domain of SARS-CoV-2. This will allow the rapid
detection of SARS-CoV-2 infections to aid in managing the current COVID-19 pandemic.
In the future, these tools could be integrated into a rapid diagnostic test for
SARS-CoV-2 and other pathogens, ultimately improving the accessibility and
sensitivity of rapid diagnostics on multiple fronts.</p>
| Identifer | oai:union.ndltd.org:purdue.edu/oai:figshare.com:article/15062847 |
| Date | 28 July 2021 |
| Creators | Kaleb M. Byers (11192533) |
| Source Sets | Purdue University |
| Detected Language | English |
| Type | Text, Thesis |
| Rights | CC BY 4.0 |
| Relation | https://figshare.com/articles/thesis/Platforms_and_Molecular_Mechanisms_for_Improving_Signal_Transduction_and_Signal_Enhancement_in_Multi-step_Point-Of-Care_Diagnostics/15062847 |
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