Determination of an efficient and reliable method for PCR detection of borrelial DNA from engorged ticks

Abstract

Since ticks have been recognized as one of the most important vectors of pathogens causing serious diseases in humans, a number of studies have focused on identifying the pathogen composition as well as transmission and infection mechanisms. Although a plethora of detection methods is available today, PCR-based approach is regarded as the most sensitive and rapid. However, common challenges in molecular analyses conducted on ticks are weak amplification results because of present inhibitors, either from a mammalian bloodmeal or a male DNA in female reproductive organs. Present study aimed to evaluate which body part of an engorged tick is the most preferable as a starting material in DNA extraction for molecular detection of Borrelia burgdorferi sensu lato, causative agent of Lyme borreliosis. We analyzed 58 Ixodes ricinus ticks removed from patients in The Center for Emergency Medical Assistance of the Sarajevo Canton, Bosnia and Herzegovina. Our findings suggest using the anterior half of semi-engorged and fully-engorged ticks for DNA extraction with the purpose of Borrelia detection.