Abstract
Assay sensitivity can be a limiting factor in the use of PCR as a tool for the detection of tick-borne pathogens in blood. We evaluated the performance of Tick-borne disease Capture Sequencing Assay (TBDCapSeq), a capture sequencing assay targeting tick-borne agents, to test 158 whole blood specimens obtained from the Lyme Disease Biobank. These included samples from 98 individuals with signs and symptoms of acute Lyme disease, 25 healthy individuals residing in Lyme disease endemic areas, and 35 samples collected from patients admitted to the Massachusetts General Hospital or referred to the infectious disease clinic. Compared to PCR, TBDCapSeq had better sensitivity and could identify infections with a wider range of tick-borne agents. TBDCapSeq identified a higher rate of samples positive for Borrelia burgdorferi (8 vs. 1 by PCR) and Babesia microti (26 vs. 15 by PCR). TBDCapSeq also identified previously unknown infections with Borrelia miyamotoi, Ehrlichia, and Rickettsia species. Overall, TBDCapSeq identified a pathogen in 43 samples vs. 23 using PCR, with four co-infections detected versus zero by PCR. We conclude that capture sequencing enables superior detection of tick-borne agents relative to PCR.
Highlights
Polymerase chain reaction (PCR) is the primary molecular assay used for clinical diagnosis of tick-borne diseases (TBD)
Capture sequencing results in a substantial increase in sensitivity surpassing quantitative PCR. The utility of this approach has been documented for viral and bacterial agents, and with the recent development of the TBD Capture Sequencing Assay (TBDCapSeq), we have demonstrated its potential for detection of tick-borne pathogens (Jain et al, 2021)
We identified 11 additional samples that were positive for B. microti and seven samples that were positive for B. burgdorferi by TBDCapSeq
Summary
Polymerase chain reaction (PCR) is the primary molecular assay used for clinical diagnosis of tick-borne diseases (TBD). Blood is the most frequent specimen used in molecular testing (Hu, 2018). PCR of whole blood is highly useful for diagnosis of acute anaplasmosis and babesiosis (Sanchez et al, 2016). PCR is not typically used for detection of Borrelia burgdorferi because of low pathogen burden coupled with transient spirochetemia in blood (Benach et al, 1983; Aguero-Rosenfeld et al, 2005).
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