2-tiered antibody testing for early and late Lyme disease using only an immunoglobulin G blot with the addition of a VlsE band as the second-tier test

Clin Infect Dis. 2010 Jan 1;50(1):20-6. doi: 10.1086/648674.


Background: Standard 2-tiered immunoglobulin G (IgG) testing has performed well in late Lyme disease (LD), but IgM testing early in the illness has been problematic. IgG VlsE antibody testing, by itself, improves early sensitivity, but may lower specificity. We studied whether elements of the 2 approaches could be combined to produce a second-tier IgG blot that performs well throughout the infection.

Methods: Separate serum sets from LD patients and control subjects were tested independently at 2 medical centers using whole-cell enzyme immunoassays and IgM and IgG immunoblots, with recombinant VlsE added to the IgG blots. The results from both centers were combined, and a new second-tier IgG algorithm was developed.

Results: With standard 2-tiered IgM and IgG testing, 31% of patients with active erythema migrans (stage 1), 63% of those with acute neuroborreliosis or carditis (stage 2), and 100% of those with arthritis or late neurologic involvement (stage 3) had positive results. Using new IgG criteria, in which only the VlsE band was scored as a second-tier test among patients with early LD (stage 1 or 2) and 5 of 11 IgG bands were required in those with stage 3 LD, 34% of patients with stage 1, 96% of those with stage 2, and 100% of those with stage 3 infection had positive responses. Both new and standard testing achieved 100% specificity.

Conclusions: Compared with standard IgM and IgG testing, the new IgG algorithm (with VlsE band) eliminates the need for IgM testing; it provides comparable or better sensitivity, and it maintains high specificity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Antibodies, Bacterial / biosynthesis
  • Antibodies, Bacterial / blood*
  • Antigens, Bacterial / blood*
  • Antigens, Bacterial / immunology
  • Bacterial Proteins / blood*
  • Bacterial Proteins / immunology
  • Blotting, Western / methods
  • Humans
  • Immunoenzyme Techniques / methods*
  • Immunoglobulin G / blood*
  • Immunoglobulin M / blood*
  • Lipoproteins / blood*
  • Lipoproteins / immunology
  • Lyme Disease / blood
  • Lyme Disease / diagnosis*
  • Lyme Disease / immunology
  • Massachusetts
  • ROC Curve
  • Sensitivity and Specificity
  • Serotyping / methods


  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Bacterial Proteins
  • Immunoglobulin G
  • Immunoglobulin M
  • Lipoproteins
  • VlsE protein, Borrelia burgdorferi