title: An Expanded Reverse Line Blot Hybridization Protocol for the Simultaneous Detection of Numerous Tick-Borne Pathogens in North America.
abstract: Due to an increasing diversity of bacterial pathogens known to be transmitted by hard ticks (Acari: Ixodidae) in North America, a comprehensive assay is needed to detect and differentiate among these numerous tick-borne pathogens. We describe an expanded protocol using a combination of multiplex polymerase chain reaction and reverse line blot hybridization to detect a greater diversity of infectious agents than were previously detectable. Ten novel oligonucleotide probes, either individually or in concert, enabled or enhanced identification of six Borrelia species, three Rickettsia species, and one Ehrlichia species. Simultaneous detection of these numerous tick-borne pathogens can advance surveillance efforts and improve accuracy of detection and, thus, reporting.
authors.affiliation: Department of Entomology, University of Illinois Urbana-Champaign, 505 S. Goodwin Ave., Urbana, IL 61801 (lpfreder@illinois.edu; johnny5yu@gmail.com; ballan@illinois.edu).
authors.forename: L Page
authors.initials: LP
authors.lastname: Fredericks
authors.affiliation: Interdisciplinary Program of Genetics, Texas A&M University, 2128 TAMU, College Station, TX 77843 (forgacs1@tamu.edu).
authors.forename: David
authors.initials: D
authors.lastname: Forgacs
authors.affiliation: Department of Entomology, University of Illinois Urbana-Champaign, 505 S. Goodwin Ave., Urbana, IL 61801 (lpfreder@illinois.edu; johnny5yu@gmail.com; ballan@illinois.edu).
authors.forename: Johnny
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Entomology, University of Illinois Urbana-Champaign, 505 S. Goodwin Ave., Urbana, IL 61801 (lpfreder@illinois.edu; johnny5yu@gmail.com; ballan@illinois.edu), ballan@illinois.edu.|School of Integrative Biology, University of Illinois Urbana-Champaign, 505 S. Goodwin Ave., Urbana, IL 61801, and.
authors.forename: Brian F
authors.initials: BF
authors.lastname: Allan
mesh_terms: D000818:Animals; D001096:Arachnid Vectors / Q000382:microbiology* / Q000502:physiology; D001898:Borrelia / Q000235:genetics / Q000302:isolation & purification*; D004536:Ehrlichia / Q000235:genetics / Q000302:isolation & purification*; D017403:In Situ Hybridization / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction; D009656:North America; D012281:Rickettsia / Q000235:genetics / Q000302:isolation & purification*; D017282:Tick-Borne Diseases / Q000382:microbiology* / Q000635:transmission; D013987:Ticks / Q000382:microbiology* / Q000502:physiology
keywords: Borrelia; Ehrlichia; Rickettsia; pathogen detection; tick-borne disease