# Evolution of Toxoplasma gondii Detection and Genotyping Methods

## TL;DR

This paper reviews the development of methods to detect and genotype Toxoplasma gondii, a parasite affecting humans and other hosts.

## Contribution

The study compiles and evaluates both traditional and modern molecular methods for T. gondii detection and genotyping.

## Key findings

- Molecular methods like PCR and LAMP are widely used for T. gondii detection.
- Genotyping techniques such as MLST and microsatellite analysis help assess parasite diversity.
- Newer methods like ddPCR offer improved sensitivity and specificity.

## Abstract

Toxoplasma gondii is a parasitic protozoan that was first described over 100 years ago. Since then, its presence has been confirmed in a wide range of hosts, including humans. T. gondii infections are estimated to affect approximately 30% of the human population, representing a particular hazard to pregnant women, the elderly and immunocompromised individuals. In recent decades, diagnostic methods have been developed to detect the parasite, not only in clinical specimens but also in food or environmental samples. Nowadays, in addition to the mere confirmation of the presence of the parasite, the determination of its genotype is becoming increasingly important. The assessment of T. gondii diversity enables the investigation of the parasite′s circulation in host populations, the identification of atypical isolates and the inference of virulence. The study is aimed at gathering and presenting information on methods for detecting and genotyping T. gondii, including their range of applications, advantages and limitations. In view of the rapid development of molecular methods in recent years, particular emphasis was placed on this group of techniques. Therefore, both popular methods, such as one‐step PCR and nested PCR and relatively new techniques, such as LAMP or ddPCR, are included in the discussion. The review is complemented by the presentation of the most common techniques used in T. gondii genotyping, including PCR‐RFLP, MLST and microsatellite sequence polymorphism studies.