Detection of , , , , and Powassan Virus in Ticks by a Multiplex Real-Time Reverse Transcription-PCR Assay.

ticks are implicated in transmission of , , , , and Powassan virus. We describe the establishment and implementation of the first multiplex real-time PCR assay with the capability to simultaneously detect and differentiate all five pathogens in a single reaction. The application of this assay for analysis of ticks at sites in New York and Connecticut revealed a high prevalence of in ticks from Suffolk County, NY. These findings are consistent with reports of a higher incidence of babesiosis from clinicians managing the care of patients with tick-borne diseases in this region. The understanding of pathogen prevalence is an important factor in the determination of human risks for tick-borne diseases and can help guide diagnosis and treatment. The implementation of our assay addresses a critical need in surveillance of tick-borne diseases, through generation of a comprehensive assessment of pathogen prevalence in . Our finding of a high frequency of ticks infected with in Suffolk County, NY, implicates this agent as a probable frequent cause of non-Lyme tick-borne disease in this area.