Detection of Babesia, Borrelia, Anaplasma, and Rickettsia spp. in Adult Black-Legged Ticks (Ixodes scapularis) from Pennsylvania, United States, with a Luminex Multiplex Bead Assay

Abstract

Ixodes scapularis, the black-legged tick, harbors multiple organisms and transmits several pathogens to animals and humans. To determine the presence of tick-borne microorganisms carried by I. scapularis in Pennsylvania, 299 adult I. scapularis ticks were collected from across the state and tested with a multiplex bead panel targeting 20 microorganisms. The Luminex bead-based xMAP Ò MultiFLEX Mega Tick Panel detected microorganisms in these ticks, including Anaplasma spp. (1.7%), Borrelia spp. (45.8%), Babesia spp. (16.1%), and Rickettsia spp. (22.1%) at the genera level and identified Anaplasma phagocytophilum (1.7%), Babesia microti (0.7%), Borrelia burgdorferi sensu stricto (45.5%), Borrelia miyamotoi (0.3%), and Rickettsia parkeri (0.7%) at the species level. Babesia spp. reactivity was found to be due to Ba. odocoilei, and Rickettsia spp. reactivity was mainly due to rickettsial endosymbionts.

Pentaplex real‐time PCR for differential detection of Yersinia pestis and Y. pseudotuberculosis and application for testing fleas collected during plague epizootics

“…mayonii ), Trypanosoma cruzi , and Leptospira interrogans for specificity testing (Table 3). The reason we included these species is that the transmission route of these bacteria is similar to the Yersinia species, either by fleas or other blood‐feeding vectors (Bai et al, 2017; Brook et al, 2015; Livengood, Hutchinson, Thirumalapura, & Tewari, 2020; Pawelczyk, Asman, & Solarz, 2019) or by consumption of contaminated water (Casanovas‐Massana, Pedra, Wunder, Begon, & Ko, 2018). All DNA from these non‐ Yesinia species was obtained from CDC Bacterial Diseases Branch.…”

“…Two Y. enterocolitica strains were included for specificity checking (Table 2) Leptospira interrogans for specificity testing ( Table 3). The reason we included these species is that the transmission route of these bacteria is similar to the Yersinia species, either by fleas or other bloodfeeding vectors (Bai et al, 2017;Brook et al, 2015;Livengood, TA B L E 1 Description of the genes selected for the pentaplex real-time PCR and the validation PCR, and the primers/probes sequences (Table 3) by running a pentaplex real-time PCR using the optimized primer concentrations.…”

Detection of Anaplasma phagocytophilum, Babesia odocoilei, Babesia sp., Borrelia burgdorferi Sensu Lato, and Hepatozoon canis in Ixodes scapularis Ticks Collected in Eastern Canada

“…Pertinent to I. scapularis ticks, B. odocoilei has transovarial transmission (female to eggs to larvae) [19,21] and, similarly, transstadial passage (larva to nymph or nymph to adult) [8,14,19,21]. The present epidemiological study, which is centered directly in the territorial heartland of I. scapularis and white-tailed deer, Odocoileus virginianus, was conducted in the Great Lakes basin [22][23][24].…”

Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera Anaplasma and Ehrlichia in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies

“…The assay involves in vitro amplification of a 100 bp 16S rDNA gene fragment targeting six different species of Anaplasma and Ehrlichia and is followed by the capture and detection of species-specific amplicons using pathogen-specific complementary oligonucleotide probes attached to magnetic beads. The xMAP hybridization assay offers a distinct advantage similar to several previously reported similar methods for detecting human and veterinary pathogens [56][57][58][59].…”