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12 mars 2011 6 12 /03 /mars /2011 09:59

Clin Vaccine Immunol. 2011 Mar 2.

Multiplex immunoassay for Lyme disease using VlsE1-IgG and pepC10-IgM antibodies: improving test performance through bioinformatics.

Porwancher RBHagerty CGFan JLandsberg LJohnson BJKopnitsky MSteere ACKulas KWong SJ.

Department of Medicine, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, New Jersey; Infectious Disease Consultants, PC, Mercerville, New Jersey; Department of Operations Research and Financial Engineering, Princeton University, Princeton, New Jersey; Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado; Zeus Scientific, Inc., Branchburg, New Jersey; Division of Rheumatology, Allergy, and Immunology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts; Wadsworth Center, New York State Department of Health, Albany, New York.

The Centers for Disease Control and Prevention currently recommends a 2-tier serologic approach to Lyme disease laboratory diagnosis, comprised of an initial serum enzyme immunoassay (EIA) for antibody to Borrelia burgdorferi followed by supplementary IgG and IgM Western blotting of EIA-positive or -equivocal samples.

Western blot accuracy is limited by subjective interpretation of weak-positive bands, false-positive IgM immunoblots, and low sensitivity for early disease.

We developed an objective alternative second-tier immunoassay using a multiplex microsphere system that measures VlsE1-IgG and pepC10-IgM antibodies simultaneously from the same sample.


Our study population was comprised of 79 patients with early acute Lyme disease, 82 with early convalescent, 47 with stages II or III disease, 34 post-antibiotic treatment patients, and 794 controls.


A bioinformatic technique called partial receiver-operating characteristic (ROC) regression was used to combine individual antibody levels into a single diagnostic score with a single cutoff; this technique enhances test performance when high specificity is required (e.g. ≥95%).


Compared to the Western blot, the multiplex assay was equally specific (95.6%), but 20.7% more sensitive for early convalescent disease (89.0% versus 68.3%, respectively; 95% CI of difference: 12.1% to 30.9%) and 12.5% more sensitive overall (75.0% versus 62.5%, respectively; 95% CI of difference: 8.1% to 17.1%).


When used as a second-tier test, a multiplex assay for VlsE1-IgG and pepC10-IgM antibodies performed as well as or better than the Western blot for Lyme disease diagnosis. Prospective validation studies appear to be warranted.

PMID: 21367982 [PubMed - as supplied by publisher]

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