Quantification of Borrelia burgdorferi Membrane Proteins in Human Serum: A New Concept for Detection of Bacterial Infection.

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TitleQuantification of Borrelia burgdorferi Membrane Proteins in Human Serum: A New Concept for Detection of Bacterial Infection.
Publication TypeJournal Article
Year of Publication2015
AuthorsCheung, CSF, Anderson, KW, Benitez, KYVillator, Soloski, MJ, Aucott, JN, Phinney, KW, Turko, IV
JournalAnal Chem
Volume87
Issue22
Pagination11383-8
Date Published2015 Nov 17
ISSN1520-6882
KeywordsBorrelia burgdorferi, Humans, Lyme Disease, Membrane Proteins, Sensitivity and Specificity
Abstract

The Borrelia burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. The low abundance of bacterial proteins in human serum during infection imposes a challenge for early proteomic detection of Lyme disease. To address this challenge, we propose to detect membrane proteins released from bacteria due to disruption of their plasma membrane triggered by the innate immune system. These membrane proteins can be separated from the bulk of serum proteins by high-speed centrifugation causing substantial sample enrichment prior to targeted protein quantification using multiple reaction monitoring mass spectrometry. This new approach was first applied to detection of B. burgdorferi membrane proteins supplemented in human serum. Our results indicated that detection of B. burgdorferi membrane proteins, which are ≈10(7) lower in abundance than major serum proteins, is feasible. Therefore, quantitative analysis was also carried out for serum samples from three patients with acute Lyme disease. We were able to demonstrate the detection of ospA, the major B. burgdorferi lipoprotein at the level of 4.0 fmol of ospA/mg of serum protein. The results confirm the concept and suggest that the proposed approach can be expanded to detect other bacterial infections in humans, particularly where existing diagnostics are unreliable.

DOI10.1021/acs.analchem.5b02803
Alternate JournalAnal. Chem.
PubMed ID26491962
PubMed Central IDPMC4809138
Grant List9999-NIST / / Intramural NIST DOC / United States
P30AR05350 / AR / NIAMS NIH HHS / United States