Quantification of Borrelia burgdorferi Membrane Proteins in Human Serum: A New Concept for Detection of Bacterial Infection.
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.
Biomolecular Measurement Division, National Institute of Standards and Technology , Gaithersburg, Maryland 20899, United States.|Institute for Bioscience and Biotechnology Research , Rockville, Maryland 20850, United States.
Crystal S F
CS
Cheung
Biomolecular Measurement Division, National Institute of Standards and Technology , Gaithersburg, Maryland 20899, United States.|Institute for Bioscience and Biotechnology Research , Rockville, Maryland 20850, United States.
Kyle W
KW
Anderson
Institute for Bioscience and Biotechnology Research , Rockville, Maryland 20850, United States.
Kenia Y Villatoro
KY
Benitez
Department of Medicine, Johns Hopkins School of Medicine , Baltimore, Maryland 21224, United States.
Mark J
MJ
Soloski
Department of Medicine, Johns Hopkins School of Medicine , Baltimore, Maryland 21224, United States.
John N
JN
Aucott
Biomolecular Measurement Division, National Institute of Standards and Technology , Gaithersburg, Maryland 20899, United States.
Karen W
KW
Phinney
Biomolecular Measurement Division, National Institute of Standards and Technology , Gaithersburg, Maryland 20899, United States.|Institute for Bioscience and Biotechnology Research , Rockville, Maryland 20850, United States.
Illarion V
IV
Turko
D025065:Borrelia burgdorferi / Q000737:chemistry*; D006801:Humans; D008193:Lyme Disease / Q000097:blood / Q000175:diagnosis*; D008565:Membrane Proteins / Q000097:blood* / Q000737:chemistry; D012680:Sensitivity and Specificity
D008565:Membrane Proteins