Isothermal Point Mutation Detection: Toward a DNA-Based Rapid Diagnostic for Sickle Cell Anemia

Abstract

Sickle cell disease (SCD) is a life-threatening inherited blood disorder that affects over 300,000 newborns globally per year, with patients in low- and middle-income countries (LMICs) disproportionately impacted. We present a novel DNA-based point-of-care diagnostic method that detects the mutation in the HBB gene responsible for forming the pathological sickle hemoglobin HbS. Using a single round of isothermal amplification, our assay selectively amplifies either HbS or normal HbA target nucleic acids, separating target and non-target amplification by over 6 minutes, and takes 20 minutes to complete. A low-cost, rapid diagnostic for SCD may enable expansion of newborn screening in LMICs.