Access to diagnostics is critical for successful treatment. Limitations of trained technicians, financial resources, and infrastructure often prevent the use of conventional diagnostics in low-resource areas. Without appropriate diagnostics, clinicians may choose to overtreat, which can harm patients’ health, waste precious resources, and contribute to public health issues, such as antimicrobial resistance. In this thesis, I describe the development of three low-cost paper-based assays for cervical cancer screening and neonatal sepsis. Current cervical cancer screening methods prioritize sensitivity over specificity, leading to overtreatment that can cause unnecessary pregnancy complications and physical harm in women who naturally resolve HPV infections. To address these limitations, I developed a HPV oncoprotein test to specifically detect progression to cervical cancer. As for neonatal sepsis, infants are often empirically treated in low-resource settings where infrastructure requirements, cost, and result turnaround time limit the use of blood culture. I developed two tests for neonatal sepsis to A) safely discontinue antibiotics in neonates being treated and B) to identify neonates at high risk of sepsis. These tests utilize the natural wicking properties of paper membranes to sequentially drive reagents for immunoassay binding reactions. By automating reagent delivery, these tests minimize hands-on time and result turnaround time to minimize training requirements and allow for timely decision-making. These tests will enable better diagnosis of cervical cancer and neonatal sepsis to reduce overtreatment at the point of care.