Desarrollo de un protocolo para la detección molecular de Toxoplasma gondii mediante ensayos qPCR en diferentes muestras biológicas

Abstract

The study titled Development of a protocol for molecular detection of Toxoplasma gondii through qPCR assays in different biological samples aims to implement a more precise and efficient diagnostic method to identify this parasite in Ecuador. Toxoplasma gondii, an intracellular protozoan from the phylum Apicomplexa, is the causative agent of toxoplasmosis, a zoonotic disease that affects 30% of the global population. It poses a significant risk, particularly for pregnant women and immunocompromised individuals. Ecuador's warm and humid climate, combined with the abundance of felines, increases the risk of environmental transmission of the parasite. Additionally, the consumption of undercooked food and contaminated water further raises the incidence of the disease. The developed protocol is based on qPCR techniques, which provide greater sensitivity and specificity compared to traditional serological methods. The study conducted an exhaustive search for unique T. gondii genetic sequences using bioinformatics tools, selecting the most suitable primers and probes. Biological samples, including blood, feces, and tissues, were collected to evaluate the protocol, and a positive control from certified strains was used. The qPCR assays demonstrated high effectiveness in detecting the parasite with low cycle threshold (Ct) values, highlighting their robustness in identifying even low concentrations of DNA. This protocol significantly contributes to improving epidemiological surveillance and public health management, especially in regions with a high parasite prevalence. Finally, the research emphasizes the need to continue strengthening molecular methods for T. gondii detection in Ecuador, promoting prevention strategies and early diagnosis in vulnerable populations.