Evaluación de la eficiencia de un ensayo LAMP para la identificación de material genético de Toxoplasma gondii en leche bovina

Abstract

Toxoplasma gondii is a zoonotic protozoan parasite, considered of great importance since it is considered a public health problem and it affects a vulnerable population due to the need to implement a safe and reliable diagnostic method. Currently, the LAMP assay has proven to be a cost-effective diagnostic test for infectious microorganisms. The objective of this study was to evaluate the efficiency of the LAMP molecular technique for the identification of genetic material of Toxoplasma gondii in milk from cows whose productive destination is the production of quesillo or cheese in an artisanal way. 100 milk samples from 100 cows were obtained, subsequently aliquots of each sample were made for storage at -80 °C and purified DNA extraction following the Ghaheri method for purification. The purified DNA was made a process control to ensure that the samples obtained are correct, this by means of a LAMP assay which is carried out using a primer kit for the amplification of Bos Taurus DNA, where an agar gel was subsequently made to amplify it with electrophoresis and culminating with the photodocumentator to observe the results. Then, a LAMP assay was performed to verify the presence or not of Toxoplasma gondii genetic material following exactly the same steps that were used in the LAMP assay for process control with the difference that a different primer kit was used so that the DNA of T. gondii can be amplified. The result of the process control was favorable, 100% of the samples obtained were positive, which confirms that the samples were extracted, transported and stored correctly. On the other hand, the LAMP assay for amplification of genetic material of T. gondii was negative in all samples. This study concluding that the LAMP assay to amplify genetic material of Toxoplasma gondii did not have positive results, this may be due to several factors such as: the climatic conditions of the place, the presence of vectors, and the survival of the parasite in the organism of the cows. Etc. Therefore, this method cannot be definitively ruled out for the diagnosis of T. gondii.