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Comparative Study
. 2025 Jun 30;20(6):e0327376.
doi: 10.1371/journal.pone.0327376. eCollection 2025.

Real-world Lyme disease testing results using modified vs standard two-tier test protocols

Yonghong Li  1 , Zhen Chen  1 , Carmen H Tong  1 , Ky Van  1 , Robert S Jones  1 , Lance A Bare  1
Affiliations
Comparative Study

Real-world Lyme disease testing results using modified vs standard two-tier test protocols

Yonghong Li et al. PLoS One. .

Abstract

The modified two-tier test (MTTT) and standard two-tier test (STTT) protocols are being used for Lyme disease serology testing in the clinic. We aimed to compare the real-world testing results of MTTT, a recently approved protocol, vs STTT, a mainstay protocol over the past decades. To this end, we obtained results of Lyme disease testing performed in a US national reference laboratory in 2022 and 2023 and constructed a matched cohort with 66,708 individuals tested using MTTT and 66,708 individuals tested using STTT. We found that, compared with STTT, MTTT identified more test positives in adults aged 18 and older and similar number of test positives in the children and adolescents. The odds ratio (95% confidence interval) of testing positive using MTTT vs STTT was 1.88 (1.79-1.98) in adults and 1.09 (0.97-1.23) in non-adults. In addition, more patients tested positive for immunoglobulin M antibody alone or positive for both immunoglobulin M and immunoglobulin G antibodies using MTTT than STTT.

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Conflict of interest statement

I have read the journal's policy and the authors of this manuscript have the following competing interests: All authors (YL, ZC, CHT, KV, RSJ, and LAB) were employed by Quest Diagnostics during the study.

Figures

Fig 1
Fig 1. Flow chart of the study cohort.
Patients tested using the modified two-tier test (MTTT) protocol and the standard two-tier test (STTT) protocol were matched by age, sex, test month and year, and state of residence.
Fig 2
Fig 2. Comparison of IgM and/or IgG positivity using the modified two-tier test (MTTT) protocol with the standard two-tier test (STTT) protocol.
Distributions of IgM and/or IgG among individuals who tested positive are shown.
See this image and copyright information in PMC

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