Direct molecular detection and genotyping of Borrelia burgdorferi from whole blood of patients with early Lyme disease

Published Date
Journal
PLoS One
Citation
PLoS One. 2012;7(5):e3682
DOI
10.1371/journal.pone.0036825
Authors
Eshoo MW
Crowder CC
Rebman AW
Rounds MA,
Matthews HE
Picuri JM
Soloski MJ
Ecker DJ
Schutzer SE
Aucott JN
Abstract

Direct molecular tests in blood for early Lyme disease can be insensitive due to low amount of circulating Borrelia burgdorferi DNA. To address this challenge, we have developed a sensitive strategy to both detect and genotype B. burgdorferi directly from whole blood collected during the initial patient visit. This strategy improved sensitivity by employing 1.25 mL of whole blood, a novel pre-enrichment of the entire specimen extract for Borrelia DNA prior to a multi-locus PCR and electrospray ionization mass spectrometry detection assay. We evaluated the assay on blood collected at the initial presentation from 21 endemic area patients who had both physician-diagnosed erythema migrans (EM) and positive two-tiered serology either at the initial visit or at a follow-up visit after three weeks of antibiotic therapy. Results of this DNA analysis showed detection of B. burgdorferi in 13 of 21 patients (62%). In most cases the new assay also provided the B. burgdorferigenotype. The combined results of our direct detection assay with initial physician visit serology resulted in the detection of early Lymedisease in 19 of 21 (90%) of patients at the initial visit. In 5 of 21 cases we demonstrate the ability to detect B. burgdorferi in early Lymedisease directly from whole blood specimens prior to seroconversion.