Serum sample containing endogenous antibodies interfering with multiple hormone immunoassays. Laboratory strategies to detect interference

Pract Lab Med. 2015 Nov 27:4:1-10. doi: 10.1016/j.plabm.2015.11.001. eCollection 2016 Apr 1.

Abstract

Objectives: Endogenous antibodies (EA) may interfere with immunoassays, causing erroneous results for hormone analyses. As (in most cases) this interference arises from the assay format and most immunoassays, even from different manufacturers, are constructed in a similar way, it is possible for a single type of EA to interfere with different immunoassays. Here we describe the case of a patient whose serum sample contains EA that interfere several hormones tests. We also discuss the strategies deployed to detect interference.

Subjects and methods: Over a period of four years, a 30-year-old man was subjected to a plethora of laboratory and imaging diagnostic procedures as a consequence of elevated hormone results, mainly of pituitary origin, which did not correlate with the overall clinical picture.

Results: Once analytical interference was suspected, the best laboratory approaches to investigate it were sample reanalysis on an alternative platform and sample incubation with antibody blocking tubes. Construction of an in-house 'nonsense' sandwich assay was also a valuable strategy to confirm interference. In contrast, serial sample dilutions were of no value in our case, while polyethylene glycol (PEG) precipitation gave inconclusive results, probably due to the use of inappropriate PEG concentrations for several of the tests assayed.

Conclusions: Clinicians and laboratorians must be aware of the drawbacks of immunometric assays, and alert to the possibility of EA interference when results do not fit the clinical pattern.

Keywords: ACTH, adrenocorticotropic hormone; Case report; EA, endogenous antibodies; EQAS, external quality assurance schemes; Endogenous antibodies; FSH, follicular stimulating hormone; HCU, Hospital Clínico Universitario “Lozano Blesa”; Immunoassay; Interference; LH, luteinising hormone; MRI, magnetic resonance imaging; PEG, polyethylene glycol; Pituitary hormones; QC, quality control; TSH, thyrotropin; fT4, free thyroxine.

Publication types

  • Case Reports