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$125,000 Award

Project Overview

Although malaria is not endemic in the United States, the UMMC laboratories routinely diagnose malaria in travelers. Malaria naïve individuals are at the highest risk for developing severe malaria, which can lead to cerebral malaria and affect multiple organ systems. Prompt and accurate species-level diagnosis is critical for providing targeted therapy to patients with malaria. This project will leverage Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology to develop a diagnostic test that allows for the highly sensitive detection of nucleic acids specific for each Plasmodium species causing human malaria infection. The team aims to implement this CRISPR-based rapid diagnostic test for use in the CLIA/CAP UMMC Clinical Laboratories of Pathology to improve the malaria detection algorithm. This technology has the potential to increase sensitivity of detecting low-parasitemia infections, improve specificity in difficult-to-identify scenarios, and decrease turnaround time to obtaining species-level identification.

Project Team

  • Nicole Putnam, PhD 
  • Feng Jiang, MD, PhD
  • Sanford Stass, MD
  • Kristin Mullins, PhD
  • Matthew Laurens, MD, MPH
  • Jonathan Jacobs

Midpoint Progress Updates – October 2023

In the initial six months of their project, the team has successfully achieved several milestones. First, specific guide RNAs (gRNAs) for the identification of each of the four Plasmodium species were designed. A florescence plate reader was employed in conjunction with an imaging system which demonstrated that CRISPR-Cas12a exhibits high sensitivity in detecting all four Plasmodium species with a Limit of Detection (LOD) of 1 copy/μL. Additionally, the team expanded the CRISPR-Cas12a test to include lateral flow dipsticks for result interpretation. This paper strip-based CRISPR-Cas12a test can directly reveal results with sensitivity and specificity equivalent to those obtained using an image detection system. Importantly, it does not require specialized instruments. The team is currently in the process of applying for an Institutional Review Board (IRB) protocol to allow the use of human specimens for validating laboratory results.

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