Next-Gen Intestinal Parasite Detection, Leveraging Metataxonomics for Improved Diagnosis of Intestinal Protists and Helminths

Intestinal parasites continue to pose a significant public health burden in low- and middle-income countries and are increasingly recognized in developed regions. Traditional diagnostic methods, primarily based on microscopy, remain widely used despite limitations in sensitivity and taxonomic resolution. This study employs a next-generation sequencing NGS-based metataxonomic approach, integrated with classical phylogenetic methods, to characterize intestinal parasites in rural Colombian populations. We compare its performance with conventional microscopy, focusing on both protist and geohelminth detection. Our findings demonstrate that metataxonomics outperforms microscopy in detecting Strongyloides stercoralis, while Trichuris trichiura was more effectively identified via microscopy, likely due to resistant eggshells impeding DNA extraction. The metataxonomic approach also enabled confident species- and subtype-level classification, particularly for Blastocystis and Entamoeba spp., revealing high colonization rates and frequent mixed infections. This study underscores the diagnostic power of NGS-based metataxonomics, which proved highly effective for broad-spectrum parasite detection, while also highlighting areas for methodological improvement, especially in primer design to enhance taxonomic coverage.