Abstract

ROJAS, María del Carmen; VAZQUEZ, Pablo Mauricio  and  COSTAMAGNA, Sixto Raúl. Alternativa rápida de extracción de ADN en quistes de Acanthamoeba para uso en el laboratorio de análisis clínicos. Acta bioquím. clín. latinoam. [online]. 2022, vol.56, n.2, pp.187-193. ISSN 0325-2957.

The objective of the study was to compare the DNA extraction of Acanthamoeba sp. cysts with a commercially available method and four non-commercial ones, with heat and ultrasound treatment that allows amplification by conventional polymerase chain reaction (PCR), reducing sample preparation and extraction times, such as a tool for diagnosis in the clinical laboratory. To this aim, a strain of Acanthamoeba T4 grown on non-nutrient agar was used. Plates with cysts at three different encystation times were stored at room temperature until the study was carried out. DNA was extracted with five methods that included pretreatments (thermal and ultrasound) or combinations of them. PCR was performed using specific primers JDP1/JDP2. Concentration and purity of DNA revealed statistically significant differences (p<0.0001) between methods. Method E (commercial), method A (thermal) and B (ultrasound) got the best yields in amplifying the specific fragment of Acanthamoeba sp. by conventional PCR.

Keywords : Cyst; DNA; Acanthamoeba; Heat treatment; Ultrasound; PCR.

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