Abstract

GUEVARA-VEGA, Marco; VERTEL-MORINSON, Melba  and  PATERNINA, Luis Enrique. Comparison of three DNA extraction methods from hard ticks (Acari: Ixodidae). rev.udcaactual.divulg.cient. [online]. 2019, vol.22, n.1, e1208.  Epub May 10, 2019. ISSN 0123-4226.  https://doi.org/10.31910/rudca.v22.n1.2019.1208.

Ticks are the most important group of ectoparasites in the transmission of pathogens to domestic animals and humans. Detection of those pathogens is usually performed by PCR-based methods and therefore requires the extraction of nucleic acids for the selective amplification of molecular targets. The aim of the present investigation was to compare the performance of three DNA extraction methods (salts, columns and guanidine thiocyanate) from ticks for studies of pathogen detection and molecular systematics of ticks. DNA extraction performance was measured by multiple comparisons on 30 tick samples and assessment of quality of the DNA extract through PCR amplification of 16S mitochondrial ticks gene. The presence of Rickettsia, Ehrlichia, Anaplasma and Babesia were also evaluated. The guanidine thiocyanate was the method with highest performance (mdnR= 160ng/uL), then columns (mdnR= 4.7ng/uL) and finally salting out method (mdnR= 1.6ng/uL). Although there were statistical differences of performance among DNA extraction protocols, there were no differences regarding the success of PCR amplification according to the extraction method (p = 0.1173). No rickettsial pathogens or piroplasms were detected in any of the DNA extracts evaluated. Considering the cost/benefit ratio of the three methods, the use of the salting out method can facilitate the massification of studies on tick-borne pathogen in low-budget laboratories.

Keywords : pathogens; PCR; molecular systematics; ectoparasites; disease vectors.

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