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Blots are not a lab favorite

  • Immunoblots can be difficult - difficult to run, difficult to interpret and prone to intra-lab and inter-lab variability.
  • For 25 years, labs have followed the Standard Two-Tiered Testing (STTT) algorithm, relying on the results of immunoblots to provide additional supportive evidence of infection.
  • “The overall sensitivity of any two-tiered algorithm is equal to its least sensitive component, and therefore the Western blot component reduces the overall sensitivity of the current 2-tiered algorithm for the diagnosis of early Lyme disease.”1

Key blot problems:

  • Low sensitivity in early disease
  • Time and technical burden
  • Subjectivity interpreting test results
  • False positives
  • Reporting delays, waiting days for results
  • Messy, cumbersome, complex
  • Sending out is costly and eats into revenue

How do you deal with the gap in detecting 30+ days IgM+/IgG- Lyme positive samples?

The gap: 30+ days IgM+ / IgG-

“According to the CDC criteria, a positive second-tier IgM Western blot result can be used to support the diagnosis only in persons with a duration of illness of < 1 month2. This caveat was included to reduce the number of false-positive results, because patients were expected to have positive IgG antibody responses after 1 month of illness… it may take up to 2 months before they have IgG responses against 5 antigens, as required for a positive IgG blot result3. In the current study, 7 of 26 patients with stage 2 LD had only positive IgM responses 5-8 weeks after disease onset; therefore, they were classified as seronegative by standard 2-tiered testing, reducing the clinical sensitivity of the standard algorithm. In contrast, the proposed 2-EIA algorithm is meant to be applied to any patient, regardless of duration of illness. This modification improves sensitivity in stage 2 LD, compared with standard 2-tiered testing, and it obviates the need to pinpoint the exact duration of illness, which can be difficult.”4

Filling the gap: ZEUS Borrelia MTTT®

With ZEUS Borrelia MTTT, there is no time limitation surrounding the second-tier EIA result, addressing the variability observed in individual IgG/IgM responses and supporting accurate diagnoses.


Now, with ZEUS Borrelia MTTT you can eliminate blots altogether!

ZEUS Borrelia MTTT algorithms…

    • Are the new algorithms for the detection of Lyme disease
      • Replaces second-tier immunoblot with a more sensitive and comparably specific ZEUS ELISA
    • Deliver higher sensitivity over STTT using ZEUS ELISA ™ assays
      • Detects up to 30% more acute Lyme disease cases compared with STTT*
    • Enable a streamlined workflow for shorter turn around time
      • Saves labs time and report results to the physician faster


Replace the immunoblot with an ELISA assay and eliminate the technical complexities and inherent variability. With ZEUS Borrelia MTTT, simple and flexible first-tier and second-tier tests can all be performed right in your laboratory.






  1. Branda JA, Body BA, Boyle J, Branson BM, Dattwyler RJ, Fikrig E, Gerald NJ, Gomes-Solecki M, Kintrup M, Ledizet M, Levin AE, Lewinski M, Liotta LA, Marques A, Mead PS, Mongodin EF, Pillai S, Rao P, Robinson WH, Roth KM, Schriefer ME, Slezak T, Snyder J, Steere AC, Witkowski J, Wong SJ, Schutzer SE.  Advances in Serodiagnostic Testing for Lyme Disease Are at Hand. Clin Infect Dis 2018 Mar 19;66 7:1133-1139.
  2. Recommendations for test performance and interpretation from the Second National Conference on Serologic Diagnosis of Lyme Disease, MMWR Morb Mortal Wkly Rep, 1995, vol. 44 (pg. 590-1)
  3. Branda JA,  Aguero-Rosenfeld ME,  Ferraro MJ,  Johnson BJ,  Wormser GP,  Steere AC. 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, vol. 50 (pg. 20-6)
  4. Branda, J.A., Linskey, K., Kim, Y.A., Steere, A.C., and Ferraro, M.J. Two-Tiered Antibody Testing for Lyme Disease With Use of 2 Enzyme Immunoassays, a Whole-Cell Sonicate Enzyme Immunoassay Followed by a VlsE C6 Peptide Immunoassay Clin Infect Dis. 2011 Sep;53(6):541-7.


* Data on file, ZEUS Scientific