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Published on: June 16, 2023
Macrocalcitonin Is a Novel Pitfall in the Routine of Serum Calcitonin Immunoassay
Thalita G Alves1, Teresa S Kasamatsu1, Ji H Yang1
1Thyroid Disease Center and Laboratory of Molecular and Translational Endocrinology (T.G.A., T.S.K., J.H.Y., I.S.K., S.C.L., C.P.C., M.R.D.d.S., R.M.B.M., J.G.H.V., J.R.M.M.), Division of Endocrinology, Department of Medicine, Escola Paulista de Medicina, Universidade Federal de São Paulo, 04039-032 São Paulo, SP, Brazil; and Molecular Biology Division (T.G.A., M.C.Z.M., A.M., J.R.M.M.), Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo, 04044-020 São Paulo, SP, Brazil.
Context:
Calcitonin (CT) is a sensitive marker of medullary thyroid carcinoma (MTC) and is used for primary diagnosis and follow-up after thyroidectomy. However, persistently elevated CT is observed even after complete surgical removal without evidence of a recurrent or persistent tumor.
Objective:
To investigate the presence of assay interference in the serum CT of MTC patients who are apparently without a structural disease.
Patients And Methods:
We studied three index MTC cases for CT assay interference and 14 patients with metastatic MTC. The CT level was measured using an immunofluorometric assay. Screening for assay interference was performed by determination of CT levels before and after serum treatment with polyethylene glycol. Additionally, samples were analyzed by chromatography on ultra-performance liquid chromatography and protein A-Sepharose.
Results:
Patients with biochemical and structural disease showed CT mean recovery of 84.1% after polyethylene glycol treatment, whereas patients suspected of interference showed recovery from 2-7%. The elution profile on UPLC showed that the immunometric CT from these three patients behaved like a high molecular mass aggregate (>300 kDa). Additionally, when these samples were applied to the protein A-Sepharose, CT immunoreactivity was retained on the column and was only released after lowering the pH.
Conclusions:
For the first time, our results show the presence of a novel pitfall in the CT immunoassay: "macrocalcitonin." Its etiology, frequency, and meaning remain to be defined, but its recognition is of interest and can help clinicians avoid unnecessary diagnostic investigations and treatment during the follow-up of MTC.
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