Development and Evaluation of a Multiple Detection System for Diagnosing Malaria and Other Blood Parasitic Diseases in Blood and Stool Samples from Humans and Non-Human Primates

Background/Objectives: Many Tropical diseases such as Malaria, Chagas, Human African Trypanosomiasis, and Lymphatic filariasis coexist in endemic countries, affecting more than 1 billion people worldwide, and are recognized as major global vector-borne diseases. Tackle this disease lies in a correct diagnosis, sensitive, specific, and fast. This study aims to describe and validate a new highly sensitive and specific multiple-analysis system that can effectively detect numerous etiological agents in a single test. Methods: A total of 230 human blood samples were evaluated retrospectively for parasite characterization and a total of 58 stool samples from Non-Human Primates (NHP). Results: The analytical specificity of the presented method was 100%, with no unspecific amplifications or cross-reactions with other blood parasitic diseases. The detection limit obtained was between 0.6 to 3.01 parasites/µl for Plasmodium species, 1,8 parasite/ml for Trypanosomatidae, and 2 microfilariae/ml in the case of Filariae. The sensitivity, specificity, predictive values, and kappa coefficient reached almost 100%, except for the sensitivity for Filariae, which dropped to 93,9%, and its NPV to 89.5%. The operational features described a turnaround and a hands-on time notably shorter than the compared methods with a lower cost per essay. Conclusions: This work presents a cost-effective and highly sensitive multiplexed tool (RT-PCR-bp) capable of performing a simultaneous detection for blood parasitic diseases using specific fluorescence probes, enabling the diagnosis of low parasite loads and co-infections.