An innovative immunoassay for accurate aldosterone quantification: overcoming low-level inaccuracy and renal dysfunction-associated interference
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Kaijuan Wang
,
Hongying Cong
Abstract
Objectives
Accurate quantification of aldosterone is critical for screening and diagnosing primary aldosteronism (PA). Current competitive chemiluminescence immunoassays (cCLIA) overestimate plasma aldosterone concentration (PAC) compared to liquid chromatography-tandem mass spectrometry (LC-MS/MS). However, LC-MS/MS is technically demanding and time-consuming, limiting its widespread clinical utility. Therefore, a novel two-step sandwich chemiluminescence immunoassay (sCLIA) for accurate quantification of PAC was systematically evaluated.
Methods
Precision, trueness, linear range, and maximum dilution factor of the new immunoassay were comprehensively validated. In a multicenter study involving 2,696 samples from seven Chinese centers, PAC measurements were performed in parallel using sCLIA, cCLIA, and LC-MS/MS. The study specifically focused on evaluating the assay’s performance at low aldosterone concentrations and in patients with chronic kidney disease (CKD), investigating potential interference from renal impairment by comparing the consistency between immunoassays and LC-MS/MS results across different CKD stages.
Results
The sCLIA exhibited excellent analytical performance for PAC measurement, with intra-assay imprecision <4.64 % and bias <5.71 % against certificated reference materials. The assay exhibited a wide reportable range (30–100,000 ng/L) with a limit of quantification at 30 ng/L and dilution capability ≥50-fold. Compared to cCLIA, sCLIA showed superior agreement with LC-MS/MS, particularly at low PAC concentrations (<110 ng/L) and in subjects with reduced renal function (eGFR<60 mL/min/1.73 m2).
Conclusions
This novel sCLIA method exhibited excellent analytical performance, combining the practical advantages of immunoassays with LC-MS/MS accuracy, thereby offering an ideal solution for large-scale primary aldosteronism screening in clinical practice.
Funding source: CAMS Innovation Fund for Medical Sciences (CIFMS)
Award Identifier / Grant number: 2023-I2M-C&T-B-062
Funding source: Noncommunicable Chronic Diseases-National Science and Technology Major Project
Award Identifier / Grant number: 2024ZD0533201
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Research ethics: The research related to human use has been complied with all the relevant national regulations, institutional policies and in accordance the tenets of the Helsinki Declaration (as revised in 2013), and has been approved by the Ethics Committee of Fuwai Hospital (2,023–2,260) and the other six participating hospitals.
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Informed consent: Informed consent was obtained from all individuals included in this study.
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Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
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Use of Large Language Models, AI and Machine Learning Tools: None declared.
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Conflict of interest: The authors state no conflict of interest.
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Research funding: This study was funded by the CAMS Innovation Fund for Medical Sciences (CIFMS, 2023-I2M-C&T-B-062), Noncommunicable Chronic Diseases-National Science and Technology Major Project (2024ZD0533200).
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Data availability: The data that support the findings of this study are available from the corresponding author[Zhou Zhou], upon reasonable request.
References
1. Mancia, G, Kreutz, R, Brunström, M, Burnier, M, Grassi, G, Januszewicz, A, et al.. 2023 ESH guidelines for the management of arterial hypertension the task force for the management of arterial hypertension of the European society of hypertension: endorsed by the international society of hypertension (ISH) and the European renal association (ERA). J Hypertens 2023;41:1874–2071.10.1097/HJH.0000000000003480Suche in Google Scholar PubMed
2. Monticone, S, D’Ascenzo, F, Moretti, C, Williams, TA, Veglio, F, Gaita, F, et al.. Cardiovascular events and target organ damage in primary aldosteronism compared with essential hypertension: a systematic review and meta-analysis. Lancet Diabetes Endocrinol 2018;6:41–50.10.1016/S2213-8587(17)30319-4Suche in Google Scholar PubMed
3. Reincke, M, Bancos, I, Mulatero, P, Scholl, UI, Stowasser, M, Williams, TA. Diagnosis and treatment of primary aldosteronism. Lancet Diabetes Endocrinol 2021;9:876–92.10.1016/S2213-8587(21)00210-2Suche in Google Scholar PubMed
4. Mulatero, P, Monticone, S, Deinum, J, Amar, L, Prejbisz, A, Zennaro, M-C, et al.. Genetics, prevalence, screening and confirmation of primary aldosteronism: a position statement and consensus of the working group on endocrine hypertension of the European society of hypertension. J Hypertens 2020;38:1919–28.10.1097/HJH.0000000000002510Suche in Google Scholar PubMed
5. Funder, JW, Carey, RM, Mantero, F, Murad, MH, Reincke, M, Shibata, H, et al.. The management of primary aldosteronism: case detection, diagnosis, and treatment: an endocrine society clinical practice guideline. J Clin Endocrinol Metab 2016;101:1889–916.10.1210/jc.2015-4061Suche in Google Scholar PubMed
6. Zhou, W, Deng, Y, Ma, W, Zhao, H, Wang, K, Zhang, Q, et al.. Insight into the status of plasma renin and aldosterone measurement: findings from 526 clinical laboratories in China. Clin Chem Lab Med CCLM 2024;62:2233–41.10.1515/cclm-2024-0373Suche in Google Scholar PubMed
7. Zeng, Q, Li, J, Yang, Y, He, Y, Song, Y, Hu, J, et al.. Comparison of a chemiluminescence immunoassay with LC-MS/MS in the determination of the plasma aldosterone concentration in patients with impaired renal function. Steroids 2025;213:109540.10.1016/j.steroids.2024.109540Suche in Google Scholar PubMed
8. Lam, L, Chiu, WW, Davidson, JS. Overestimation of aldosterone by immunoassay in renal impairment. Clin Chem 2016;62:890–1.10.1373/clinchem.2016.255737Suche in Google Scholar PubMed
9. Handelsman, DJ, Wartofsky, L. Requirement for mass spectrometry sex steroid assays in the journal of clinical endocrinology and metabolism. J Clin Endocrinol Metab 2013;98:3971–3.10.1210/jc.2013-3375Suche in Google Scholar PubMed
10. Constantinescu, G, Bidlingmaier, M, Gruber, M, Peitzsch, M, Poitz, DM, Van Herwaarden, AE, et al.. Mass spectrometry reveals misdiagnosis of primary aldosteronism with scheduling for adrenalectomy due to immunoassay interference. Clin Chim Acta 2020;507:98–103.10.1016/j.cca.2020.04.019Suche in Google Scholar PubMed
11. Yin, Y, Ma, C, Yu, S, Liu, W, Wang, D, You, T, et al.. Comparison of three different chemiluminescence assays and a rapid liquid chromatography tandem mass spectrometry method for measuring serum aldosterone. Clin Chem Lab Med CCLM 2019;58:95–102.10.1515/cclm-2019-0706Suche in Google Scholar PubMed
12. Naruse, M, Katabami, T, Shibata, H, Sone, M, Takahashi, K, Tanabe, A, et al.. Japan endocrine society clinical practice guideline for the diagnosis and management of primary aldosteronism 2021. Endocr J 2022;69:327–59.10.1507/endocrj.EJ21-0508Suche in Google Scholar PubMed
13. McEvoy, JW, McCarthy, CP, Bruno, RM, Brouwers, S, Canavan, MD, Ceconi, C, et al.. 2024 ESC Guidelines for the management of elevated blood pressure and hypertension. Eur Heart J 2024;45:3912–4018.10.1093/ehjcvp/pvae084Suche in Google Scholar PubMed PubMed Central
14. Deng, L, Xiong, Z, Li, H, Lei, X, Cheng, L. Analytical validation and investigation on reference intervals of aldosterone and renin in Chinese Han population by using fully automated chemiluminescence immunoassays. Clin Biochem 2018;56:89–94.10.1016/j.clinbiochem.2018.04.016Suche in Google Scholar PubMed
15. Fortunato, A, Prontera, C, Masotti, S, Franzini, M, Marchetti, C, Giovannini, S, et al.. State of the art of aldosterone immunoassays. A multicenter collaborative study on the behalf of the cardiovascular biomarkers study group of the Italian section of European society of ligand assay (ELAS) and società italiana di biochimica clinica (SIBIOC). Clin Chim Acta Int J Clin Chem 2015;444:106–12.10.1016/j.cca.2015.01.028Suche in Google Scholar PubMed
16. Ozeki, Y, Tanimura, Y, Nagai, S, Nomura, T, Kinoshita, M, Shibuta, K, et al.. Development of a new chemiluminescent enzyme immunoassay using a two-step sandwich method for measuring aldosterone concentrations. Diagnostics 2021;11:433.10.3390/diagnostics11030433Suche in Google Scholar PubMed PubMed Central
17. Eisenhofer, G, Kurlbaum, M, Peitzsch, M, Constantinescu, G, Remde, H, Schulze, M, et al.. The saline infusion test for primary aldosteronism: implications of immunoassay inaccuracy. J Clin Endocrinol Metab 2022;107:e2027–36.10.1210/clinem/dgab924Suche in Google Scholar PubMed PubMed Central
18. Wu, V-C, Yang, S-Y, Lin, J-W, Cheng, B-W, Kuo, C-C, Tsai, C-T, et al.. Kidney impairment in primary aldosteronism. Clin Chim Acta 2011;412:1319–25.10.1016/j.cca.2011.02.018Suche in Google Scholar PubMed
19. Kawashima, A, Sone, M, Inagaki, N, Takeda, Y, Itoh, H, Kurihara, I, et al.. Renal impairment is closely associated with plasma aldosterone concentration in patients with primary aldosteronism. Eur J Endocrinol 2019;181:339–50.10.1530/EJE-19-0047Suche in Google Scholar PubMed
Supplementary Material
This article contains supplementary material (https://doi.org/10.1515/cclm-2025-0743).
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