Interdisciplinary Initiatives Program Round 10 - 2020
Juan G. Santiago, Mechanical Engineering
Niaz Banaei, Pathology and Medicine (Infectious Diseases)
Tuberculosis (TB) is one of the most prevalent and deadly infectious diseases in the world. Tuberculosis is especially challenging to diagnose in children who have difficulty producing sputum, as well as in HIV-infected patients and extrapulmonary TB cases, and often invasive sampling is required given current techniques. Thus, there is a strong need for a noninvasive modality for TB diagnosis. Infectious agents such as the bacterium causing tuberculosis (Mycobacterium tuberculosis) introduce traces of their nucleic acids (e.g. DNA and RNA) into the blood of person which they infect. These nucleic acids in the blood are called circulating DNA/RNA, and they can be detected and identified as a way of diagnosing the disease.
Our goal is to enable sensitive and rapid detection of such nucleic acids by combining our microfluidic technology with a new enzymatic assay which can sensitively and specifically detect nucleic acids. Our microfluidic technology uses electric fields in a process called isotachophoresis (ITP) to focus DNA and RNA in microchannels together with an enzyme system called CRISPR-Cas12a. This focusing increases the enzyme activity by several orders of magnitude, dramatically improving speed and sensitivity of the detection. We will integrate this assay into a portable and automated detection device. The device will include a microfluidic chip, miniature laser and photodiode detector, electronics, and a microprocessor to control and read the assay using a phone or laptop. Importantly, our technology is easily reconfigurable to other molecular targets. For example, with a simple change in one synthetic RNA sequence, our method can also be used for a rapid and sensitive diagnostic tool applicable to several other pathogens of global health importance, including the COVID-19 virus. We believe our technology would strongly impact the emerging field of personalized medicine in which rapid identification of specific pathogens from blood, urine, or saliva enables rapid response to outbreaks, informed choice of treatments, and timely diagnosis for critically ill patients. The portable system and methods we will develop can enable field deployable solutions, which are very valuable during global health crises.