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

Mark W. Eshoo, Christopher C. Crowder, Alison W. Rebman, Megan A. Rounds, Heather E. Matthews, John M. Picuri, Mark J Soloski, David J. Ecker, Steven E. Schutzer, John Aucott

Research output: Contribution to journalArticle

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. burgdorferi genotype. The combined results of our direct detection assay with initial physician visit serology resulted in the detection of early Lyme disease 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 Lyme disease directly from whole blood specimens prior to seroconversion.

Original languageEnglish (US)
Article numbere36825
JournalPLoS One
Volume7
Issue number5
DOIs
StatePublished - May 8 2012

Fingerprint

Borrelia burgdorferi
Lyme disease
Lyme Disease
genotyping
Blood
Assays
blood
assays
Serology
physicians
DNA
Genotype
Physicians
Borrelia
Electrospray ionization
erythema
hematologic tests
Electrospray Ionization Mass Spectrometry
genotype
Hematologic Tests

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Direct molecular detection and genotyping of Borrelia burgdorferi from whole blood of patients with early Lyme disease. / Eshoo, Mark W.; Crowder, Christopher C.; Rebman, Alison W.; Rounds, Megan A.; Matthews, Heather E.; Picuri, John M.; Soloski, Mark J; Ecker, David J.; Schutzer, Steven E.; Aucott, John.

In: PLoS One, Vol. 7, No. 5, e36825, 08.05.2012.

Research output: Contribution to journalArticle

Eshoo, MW, Crowder, CC, Rebman, AW, Rounds, MA, Matthews, HE, Picuri, JM, Soloski, MJ, Ecker, DJ, Schutzer, SE & Aucott, J 2012, 'Direct molecular detection and genotyping of Borrelia burgdorferi from whole blood of patients with early Lyme disease', PLoS One, vol. 7, no. 5, e36825. https://doi.org/10.1371/journal.pone.0036825
Eshoo, Mark W. ; Crowder, Christopher C. ; Rebman, Alison W. ; Rounds, Megan A. ; Matthews, Heather E. ; Picuri, John M. ; Soloski, Mark J ; Ecker, David J. ; Schutzer, Steven E. ; Aucott, John. / Direct molecular detection and genotyping of Borrelia burgdorferi from whole blood of patients with early Lyme disease. In: PLoS One. 2012 ; Vol. 7, No. 5.
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