Toxoplasmosis is a parasitic zoonotic disease widely distributed worldwide and is caused by the intracellular parasite Toxoplasma gondii. The definitive host of T. gondii is the domestic cat and the entire cat family, in which the sexual stages of the parasite develop. T. gondii can also infect a wide range of intermediate hosts, affecting most warm-blooded animals including humans. In humans, toxoplasmosis is usually asymptomatic in healthy individuals, but can develop lymphadenopathy and nonspecific symptomatology or even be fatal in infants with congenital toxoplasmosis and in immunocompromised patients. Transmission to humans is mainly through food, especially by eating undercooked meat or meat contaminated with tissue cysts. This has led to various public health organizations worldwide monitoring programs on T. gondii in animals intended for human consumption, especially in meat samples. One of the techniques employed in the laboratory is that based on the polymerase chain reaction and some of its variants, which have proven to be valuable tools for the detection of T. gondii in tissues for human consumption and many other types of biological samples. The development of different strategies for the molecular detection of T. gondii has led to the identification and quantification methodologies varying widely among laboratories. Therefore, this chapter reviews the main methods of extraction, purification, detection and quantification of T. gondii DNA in tissue samples from different species destined for human consumption.