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Licensed Unlicensed Requires Authentication Published by De Gruyter November 29, 2019

Interference of anti-streptavidin antibodies in immunoassays: a very rare phenomenon or a more common finding?

  • Nick Verougstraete , Mario Berth ORCID logo , Mario Vaneechoutte , Joris Delanghe EMAIL logo and Nico Callewaert



Anti-streptavidin antibodies (ASA) may cause analytical interference on certain immunoassay platforms. Streptavidin is purified from the non-pathogenic Streptomyces avidinii soil bacterium. In contrast to interference with biotin, ASA interference is supposed to be much rarer. In-depth studies on this topic are lacking. Therefore, we carried out an analysis toward the prevalence and the possible underlying cause of this interference.


Anti-streptavidin (AS)-immunoglobulin G (IgG) and AS-IgM concentrations were determined on multiple samples from two patients with ASA interference and on 500 random samples. On a subset of 100 samples, thyroid-stimulating hormone (TSH) was measured on a Cobas analyzer before and after performing a neutralization protocol which removes ASA. The relationship between the ratio of TSH after neutralization/TSH before neutralization and the ASA concentration was evaluated. Subsequently, an extract of S. avidinii colonies was analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting.


A positive correlation between AS-IgM concentrations and TSH ratio was obtained. Eight samples out of 500 exceeded the calculated AS-IgM cut-off value. In comparison to the AS-IgM concentrations in the population, titers from the two described cases clearly stood out. The isolated cases represent the end of a broader spectrum as there is a continuum of AS-IgM reactivity in the general population. We could not observe any differences in the immunoblot patterns between the cases and controls, which may indicate the general presence of ASA in the population.


Interference due to ASA is more prevalent than initially thought and is caused by IgM antibodies.

Corresponding author: Prof. Dr. Joris, Department of Diagnostic Sciences, Ghent University, C. Heymanslaan 10, 9000 Ghent, Belgium, Phone: +3293322956, Fax: +3293324985
aJoris Delanghe and Nico Callewaert contributed equally to this work.


We would like to thank Elke Lecocq, Geert Esprit and Sofie Willaert for their skillful assistance.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in writing of the report; or in the decision to submit the report for publication.


1. American Association for Clinical Chemistry. AACC guidance document on biotin interference in laboratory tests. Available at: Accessed Aug 7 2019.Search in Google Scholar

2. Elston MS, Sehgal S, Du Toit S, Yarndley T, Conaglen JV. Factitious Graves’ disease due to biotin immunoassay interference – a case and review of the literature. J Clin Endocrinol Metab 2016;101:3251–5.10.1210/jc.2016-1971Search in Google Scholar PubMed

3. Piketty ML, Polak M, Flechtner I, Gonzales-Briceño L, Souberbielle JC. False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences. Clin Chem Lab Med 2017;55:780–8.10.1515/cclm-2016-0606Search in Google Scholar PubMed

4. Li J, Wagar EA, Meng QH. Comprehensive assessment of biotin interference in immunoassays. Clin Chim Acta 2018;487:293–8.10.1016/j.cca.2018.10.013Search in Google Scholar PubMed

5. Favresse J, Lardinois B, Nassogne MC, Pneumont V, Maiter D, Gruson D. Anti-streptavidin antibodies mimicking heterophilic antibodies in thyroid functions tests. Clin Chem Lab Med 2018;56:160–3.10.1515/cclm-2017-1027Search in Google Scholar PubMed

6. Rulander NJ, Cardamone D, Senior M, Snyder PJ, Master SR. Interference from anti-streptavidin antibody. Arch Pathol Lab Med 2013;137:1141–6.10.5858/arpa.2012-0270-CRSearch in Google Scholar PubMed

7. Peltier L, Massart C, Moineau MP, Delhostal A, Roudaut N. Anti-streptavidin interferences in Roche thyroid immunoassays: a case report. Clin Chem Lab Med 2016;54:11–4.10.1515/cclm-2015-0350Search in Google Scholar PubMed

8. Bayart JL, Favresse J, Melnik E, Lardinois B, Fillée C, Maiter D, et al. Erroneous thyroid and steroid hormones profile due to anti-streptavidin antibodies. Clin Chem Lab Med 2019;57:e255–8.10.1515/cclm-2018-1355Search in Google Scholar PubMed

9. Raverot V, Bordeau E, Periot C, Perrin P, Chardon L, Plotton I, et al. Letter to the editor: a case of laboratory-generated “thyroid dysfunction”. Ann Endocrinol 2019;80:140–1.10.1016/j.ando.2018.10.001Search in Google Scholar PubMed

10. Piketty ML, Prie D, Sedel F, Bernard D, Hercend C, Chanson P, et al. High-dose biotin therapy leading to false biochemical endocrine profiles: validation of a simple method to overcome biotin interference. Clin Chem Lab Med 2017;55:817–25.10.1515/cclm-2016-1183Search in Google Scholar PubMed

11. Berth M, Willaert S, De Ridder C. Anti-streptavidin IgG antibody interference in anti-cyclic citrullinated peptide (CCP) IgG antibody assays is a rare but important cause of false-positive anti-CCP results. Clin Chem Lab Med 2018;56:1263–8.10.1515/cclm-2017-1153Search in Google Scholar PubMed

12. Sànchez-Carbayo M, Mauri M, Alfayate R, Miralles C, Soria F. Analytical and clinical evaluation of TSH and thyroid hormones by electrochemiluminescent immunoassays. Clin Biochem 1999;32:395–403.10.1016/S0009-9120(99)00032-6Search in Google Scholar

13. Martel J, Després N, Ahnadi CE, Lachance JF, Monticello JE, Fink G, et al. Comparative multicentre study of a panel thyroid tests using different automated immunoassay platforms and specimens at high risk of antibody interference. Clin Chem Lab Med 2000;38:785–93.10.1515/CCLM.2000.112Search in Google Scholar PubMed

14. Quinn FA, Scopp R, Lach A, Drake C, Mo M, Albright J, et al. A comparison of different sample matrices for evaluating functional sensitivity, imprecision, and dilution linearity of the Abbott Architect i2000 TSH assay. Clin Chem Lab Med 2002;40:709–12.10.1515/CCLM.2002.122Search in Google Scholar PubMed

15. Bhattacharya CG. A simple method of resolution of a distribution into Gaussian components. Biometrics 1967;23:115–35.10.2307/2528285Search in Google Scholar

16. Naus AJ, Borst A, Kuppens PS. The use of patient data for the calculation of reference values for some haematological parameters. J Clin Chem Clin Biochem 1980;18:621–5.10.1515/cclm.1980.18.10.621Search in Google Scholar PubMed

17. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–5.10.1038/227680a0Search in Google Scholar PubMed

18. Van Dam A, Lee VH, Dunbar BS, Sammons DW. Principles and methods for preparing samples for protein transfer. In: Dunbar BS, editor. Protein blotting, a practical approach. New York: Oxford University Press, 1994:73–120.Search in Google Scholar

19. Lam L, Bagg W, Smith G, Chiu WW, Middleditch MJ, Lim JC, et al. Apparent hyperthyroidism caused by biotin-like interference from IgM anti-streptavidin antibodies. Thyroid 2018;28:1063–7.10.1089/thy.2017.0673Search in Google Scholar PubMed

20. Taylor PN, Razvi S, Pearce SH, Dayan CM. A review of the clinical consequences of variation in thyroid function within the reference range. J Clin Endocrinol Metab 2013;98:3562–71.10.1210/jc.2013-1315Search in Google Scholar PubMed

21. Fossati-Jimack L, Reininger L, Chicheportiche Y, Clynes R, Ravetch JV, Honjo T, et al. High pathogenic potential of low-affinity autoantibodies in experimental autoimmune hemolytic anemia. J Exp Med 1999;190:1689–96.10.1084/jem.190.11.1689Search in Google Scholar PubMed PubMed Central

22. Dahll LK, Haave EM, Aas FE, Dahl SR, Thorsby PM. Effect of anti-streptavidin interference when diagnosing thyroid disease using immunoassay analyses. Endocrine Abstracts 2019;63:GP181. doi:10.1530/endoabs.63.GP181.Search in Google Scholar

23. Chater KF. Recent advances in understanding Streptomyces. F1000Res 2016;5:2795.10.12688/f1000research.9534.1Search in Google Scholar PubMed PubMed Central

24. Sztefko K. Human natural antibodies and immunoassay. In: Sonntag O, Plebani M, editors. Immunodiagnostics and patient safety. Berlin: De Gruyter, 2011:43–52.10.1515/9783110249484Search in Google Scholar

25. Basic local alignment search tool. Available at: Accessed Aug 1 2019.Search in Google Scholar

Supplementary Material

The online version of this article offers supplementary material (

Received: 2019-10-14
Accepted: 2019-11-01
Published Online: 2019-11-29
Published in Print: 2020-09-25

©2020 Walter de Gruyter GmbH, Berlin/Boston

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