Molecular Detection of Helminths in Stool Samples: Methods, Challenges, and Applications

Helminth infections caused by soil-transmitted species, like Ascaris lumbricoides, Trichuris trichiura, and hookworms, affect over one billion people worldwide, yet accurate diagnosis remains challenging due to low sensitivity of microscopy in detecting eggs in stool samples, especially in low-intensity infections. Molecular diagnostics, particularly PCR-based detection of helminth DNA in stool samples, have emerged as more sensitive and specific alternatives. Here we review advances in DNA extraction methods that overcome inhibitors in stool, multiplex PCR assays, and next-generation sequencing technologies enabling species differentiation and detection of drug resistance markers. These molecular tools enhance epidemiological surveillance and inform control strategies. Despite challenges such as sample complexity and cost, ongoing improvements in molecular diagnostics hold promise for more effective helminth detection and management in clinical and field settings.