Two-Tier Lyme Disease Serology Test Results Can Vary According to the Specific First-Tier Test Used

Abstract

Background Variability in 2-tier Lyme disease test results according to the specific first-tier enzyme immunoassay (EIA) in children has not been examined rigorously. In this study, we compared paired results of clinical 2-tier Lyme disease tests to those of the C6 peptide EIA followed by supplemental immunoblotting (C6 2-tier test). Methods We performed a prospective cohort study of children aged ≥1 to ≤21 years who were undergoing evaluation for Lyme disease in the emergency department at 1 of 6 centers located in regions in which Lyme disease is endemic. The clinical first-tier test and a C6 EIA were performed on the same serum sample with supplemental immunoblotting if the first-tier test result was either positive or equivocal. We compared the results of the paired clinical and C6 2-tier Lyme disease test results using the McNemar test. Results Of the 1714 children enrolled, we collected a research serum sample from 1584 (92.4%). The clinical 2-tier EIA result was positive in 316 (19.9%) children, and the C6 2-tier test result was positive or equivocal in 295 (18.6%) children. The clinical and C6 2-tier test results disagreed more often than they would have by chance alone (P = .002). Of the 39 children with either a positive clinical or C6 2-tier test result alone, 2 children had an erythema migrans (EM) lesion, and 29 had symptoms compatible with early disseminated Lyme disease. Conclusions Two-tier Lyme disease test results differed for a substantial number of children on the basis of the specific first-tier test used. In children for whom there is a high clinical suspicion for Lyme disease and who have an initially negative test result, clinicians should consider retesting for Lyme disease.

Point-of-care serodiagnostic test for early-stage Lyme disease using a multiplexed paper-based immunoassay and machine learning

“…The second sample-pull ( (+) = 42, (−) = 54) was then entirely used for blind testing the performance of our xVFA diagnostic platform, yielding an AUC of 0.950, sensitivity of 90.5%, and specificity of 87.0% with respect to the seropositive and seronegative results, as summarized in Figure 5. This reported diagnostic performance of our xVFA platform achieved during the blind testing phase, to the best of our knowledge, exceeds previous POC tests for early-stage LD 31,42 . As a point of reference, Table S5 shows a comparison with the recently FDA-approved POC test from Quidel, which can be used as a first-tier test, detecting IgM and IgG antibodies using the C6 antigen.…”

“…For this work, samples were considered seropositive if any of the three EIA tests in the first tier had a positive or equivocal (borderline) result and the second tier had a positive result for either the IgM or IgG WB as defined by the CDC recommendation (≥ 2 of 3 bands for IgM WB, and ≥ 5 of 10 bands for IgG) 9 . Samples were also considered seropositive by MTTT guidelines, where a seropositive diagnosis was called from two positive or equivocal EIA tests along with the presence of EM, without the need for a positive WB result 16,31 . Additionally, all samples were confirmed negative for coinfections of Anaplasmosis and Babesia, both of which are infections also transmitted by the Ixodes tick and can produce similar constitutional symptoms to LD.…”

“…They can also have low sensitivity if the target antigen is mismatched with the underlying immunodominance. This paradigm is the reason that the performance of the two-tier algorithm can depend on which EIA is used in the first tier, as well as what strain of Bb the EIA test was designed against, B31 being the most common 31 .…”

A fluorescent plasmonic biochip assay for multiplex screening of diagnostic serum antibody targets in human Lyme disease

“…The STTT has been a useful diagnostic tool since its standardization by the Centers for Disease Control (CDC) in 1995 [13], but critical limitations have become increasingly apparent [16,17]. These include low sensitivity and low specificity for early disease [18,19], inability to monitor treatment progress or diagnose re-infection [20], inconsistencies across tests [21][22][23], and subjective interpretation of Western blot results [18,24]. Experts agree that new strategies for diagnosing LD are necessary to address these concerns, pointing to modified two-tier algorithms using only ELISAs [23,[25][26][27], as well as novel assays in various stages of development [28][29][30].…”

Diagnostic Performance of C6 Enzyme Immunoassay for Lyme Arthritis

“…

OBJECTIVES:In Lyme disease endemic areas, initial management of children with arthritis can be challenging because diagnostic tests take several days to return results, leading to potentially unnecessary invasive procedures. Our objective was to examine the role of the C6 peptide enzyme immunoassay (EIA) test to guide initial management.

METHODS:We enrolled children with acute arthritis undergoing evaluation for Lyme disease presenting to a participating Pedi Lyme Net emergency department (2015)(2016)(2017)(2018)(2019) and performed a C6 EIA test. We defined Lyme arthritis with a positive or equivocal C6 EIA test result followed by a positive supplemental immunoblot result and defined septic arthritis as a positive synovial fluid culture result or a positive blood culture result with synovial fluid pleocytosis.

…”