Detection of zoonotic human pathogens from Ixodes scapularis in Wisconsin

Extracted nucleic acids were also tested using real-time fluorescent resonance energy transfer (FRET) PCR assays for B. miyamotoi (GlpQ: Forward primer 5'-TCCAGAACATACCTTAGAAGC- 3'; reverse primer 5'-ATCAAATCTTTCACTGAGACTTA-3'; fluorescein-labeled probe 5'-GACAATGTTCCTATTATAATGCACGACCC-fl-3'; 5'-LC640-GAAATTGACACAACCACAAATGTTGCAC-3'), and Powassan virus (NS5: forward primer 5'-ACTAGAATGGCCATGACAGAC-3'; reverse primer 5'-TCATCTCTGGTGCACATCC-3'; fluorescein-labeled probe 5'-CACAAAGGCCCAGGAACCACAGC-fl-3'; red labeled probe 5'-LC640-GGCACCAGIGTGATCATGAGAGCIGT-3'); (WSH site only because ticks were stored in ethanol at WAL). [...]a relatively small number of ticks were analyzed and only nymphs were present in large numbers at the southeastern location (WAL). [...]inconsistent tick storage limited our ability to detect the full spectrum of pathogens. [...]this study demonstrates pathogen prevalence and coinfections in I. scapularis ticks among regions of the upper midwestern United States.