Detection of Rickettsia spp. in field-collected ticks using loop-mediated isothermal amplification (LAMP)

Cases of tick-borne rickettsial diseases are increasing in the Central United States, augmenting the need to develop new surveillance tools to detect changing disease patterns.  Loop-mediated isothermal amplification (LAMP) is a novel molecular tool which utilizes four to six primers and runs at one temperature in a simple water-bath.  Since the development of this assay, relatively few studies have utilized this novel approach to detect arthropod-borne pathogens in field-collected arthropods with most using LAMP to test blood samples from mammalian hosts.  The aim of this project was to design and validate a novel LAMP assay to detect Rickettsia spp. in field-collected ticks. All results were compared with a pan-specific PCR assay which targeted the 17 kDa gene of Rickettsia spp.  The 802 ticks tested using the two assays were collected during the summer of 2014 from various Oklahoma state parks. Preliminary results indicated the two tests correlate, signifying the LAMP assay can be further developed to differentiate rickettsial species.  These results show LAMP as a promising molecular surveillance tool which can be used effectively to detect pathogens in field-collected ticks.  This assay can then be further developed for use in resource-limited countries to assist with surveillance of tick-borne pathogens.