Collection and Characterization of Samples for Establishment of a Serum Repository for Lyme Disease Diagnostic Test Development and Evaluation

Abstract

Serological assays and a two-tiered test algorithm are recommended for laboratory confirmation of Lyme disease. In the United States, the sensitivity of two-tiered testing using commercially available serology-based assays is dependent on the stage of infection and ranges from 30% in the early localized disease stage to near 100% in late-stage disease. Other variables, including subjectivity in reading Western blots, compliance with two-tiered recommendations, use of different first-and second-tier test combinations, and use of different test samples, all contribute to variation in two-tiered test performance. The availability and use of sample sets from well-characterized Lyme disease patients and controls are needed to better assess the performance of existing tests and for development of improved assays. To address this need, the Centers for Disease Control and Prevention and the National Institutes of Health prospectively collected sera from patients at all stages of Lyme disease, as well as healthy donors and patients with look-alike diseases. Patients and healthy controls were recruited using strict inclusion and exclusion criteria. Samples from all included patients were retrospectively characterized by two-tiered testing. The results from two-tiered testing corroborated the need for novel and improved diagnostics, particularly for laboratory diagnosis of earlier stages of infection. Furthermore, the two-tiered results provide a baseline with samples from well-characterized patients that can be used in comparing the sensitivity and specificity of novel diagnostics. Panels of sera and accompanying clinical and laboratory testing results are now available to Lyme disease serological test users and researchers developing novel tests.

Lyme Borreliosis Serology: Performance of Several Commonly Used Laboratory Diagnostic Tests and a Large Resource Panel of Well-Characterized Patient Samples

“…b VlsE result was obtained using a visual read of the ViraStripe immunoblot. c These data have been reported previously (10). d No statistical differences were observed between the sensitivities of the VIDAS/C6 2-EIA MTTT and STTT algorithms (P values ranged from 0.22 to 1.00) when acute-phase early Lyme disease samples were tested (see Table S3 in the supplemental material).…”

“…c VlsE band striped onto the Borrelia B31 ViraStripe immunoblot (Viramed Biotech AG, Munich, Germany) and read visually. d The Marblot data were reported previously (10). Positive IgM immunoblot results were reported regardless of duration of illness prior to specimen collection.…”

“…We used the LSR that is a unique serum panel consisting of 471 sera that were acquired from patients with various stages of Lyme disease, as well as from patients and donors of multiple negative-control groups (10). Because the samples used in this study are available for investigators to use for novel laboratory diagnostic development, the information provided here is meant to be used as baseline data for comparative purposes.…”

“…The sera used in this study are from the CDC LSR and were previously described (10). Briefly, the LSR contains serum samples from well-documented Lyme disease patients and control groups.…”

“…In this study, we utilized samples from the recently available Lyme Serum Repository (LSR) (10) to compare the performances of STTT and MTTT algorithms. In addition, multiple first-and second-tier tests from differing commercial manufacturers were used, and their results were compiled individually.…”