Background: Recently, second-generation thyroglobulin (Tg) sandwich immunoassays have been used in clinical laboratories to measure the serum Tg levels, which is a tumor marker used to monitor postoperative patients with differentiated thyroid cancers. However, these immunoassays are often subject to Tg autoantibody (TgAb) interference. TgAb interference is inevitable for almost all Tg immunoassays, resulting in unreliable Tg measurement values of TgAb-positive samples.
Methods: To address TgAb interference, we have developed a novel immunoassay based on a fully automated chemiluminescent enzyme immunoassay system using the effective specimen-pretreatment process to inactivate TgAb in blood and evaluated its assay performance.
Results: The developed assay was traceable to BCR457 IRMM reference material with a limit of quantification of 0.03 ng/mL. The pretreatment process inactivated almost all TgAb in specimens and allowed accurate Tg measurements in TgAb-positive samples in which TgAb interference was observed using the immunoassays. Size-exclusion chromatography analysis of immunoreactive Tg molecule in a TgAb-positive serum verified disruption of the Tg-TgAb immune complex by the pretreatment process. Good correlation of Tg values in TgAb-negative specimens was observed between the new Tg immunoassay and the second-generation sandwich immunoassays. However, there were numerous discrepant samples on bias plots between the new Tg immunoassay and the second-generation sandwich immunoassays for TgAb-positive specimens.
Conclusions: This study indicates the new Tg immunoassay with the specimen-pretreatment process is both robust and free from interference by TgAb. Thus, this novel assay is superior to second-generation sandwich immunoassays and gives accurate Tg concentrations even for TgAb-positive cases.
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