Tick surveillance is becomming more complex as an increasing number of tickborne pathogens are recognized to cause human disease. Here, we describe a new PCR multiplex amplicon sequencing approach for potential use in tick surveillance programs targeting to commonly human-biting vectors, Ixodes scapularis and Ixodes pacificus. The PCR multiplex amplicon assay is using pan-PCR primers (genus) and is able to detect a broader range of tick-associated microorganisms, more effectively and detect co-infections of multiple pathogens in a single tick (including different species within the Borrelia burgdorferi sensu lato complex and differentiate between the human and non-human Anaplasma phagocytophilum), and required a smaller volume of test sample (thus preserving more sample for future testing). The addition of PCR primers that will aid in a molecular identification of ticks, is very helpfull when a morphological identification is not possible.
