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Abstract

Hookworm disease, caused by Ancylostoma duodenale and Necator americanus, is a soil-transmitted infection affecting over 700 million people worldwide. Transmission occurs through direct skin contact with contaminated soil, leading to health consequences including iron-deficiency anemia, malnutrition, and long-term developmental impacts. The CCU Disease Hunters program developed a PCR-based environmental screen to detect hookworm DNA in soil samples before transmission reaches human or animal hosts. Initiated in response to a request from All 4 Paws Animal Rescue, this project established protocols for soil collection, DNA extraction, and molecular identification of hookworm using the cytochrome c oxidase (COI) and cytochrome b (COB) genes. Four primers (HK1, HK2, HK3, and HK4) were evaluated for specificity and functionality. Primer HK3 was identified as the most functional primer set, producing bands of the expected size in soil samples, while primer HK1 was found to detect the hookworm family broadly rather than a specific species. Hookworm DNA was successfully detected in soil samples collected from All 4 Paws Animal Rescue, demonstrating that an environmental PCR screen can serve as a community surveillance tool. This prevention-focused approach offers a cost-effective strategy for monitoring high-risk areas and guiding proactive intervention to reduce disease transmission.

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