Abstract
Background
Although high-dose biotin interference is now considered in automated immunoassays, it has not yet been detected in manually studied research kits, especially with enzyme-linked immunosorbent assay (ELISA), and the possibility of biotin interference in these kits has not been demonstrated. The aims of our study were to determine the effects of biotin interference on various parameters, including leptin, leptin receptor (LEPR), ghrelin, acylated ghrelin, deacylated ghrelin, ghrelin receptor (GHSR), kisspeptin (KISS1), kisspeptin receptor (KISS1R), preptin, peroxisome proliferator activated receptor gamma (PPAR γ), nod-like receptor pyrin domain-containing 3 (NLRP3) and interleukin-18 (IL-18), which contribute to energy homeostasis in healthy and obese children.
Methods
Serum pools were prepared from healthy and obese individuals, and biotin concentrations in samples containing different amounts of biotin were measured via sandwich and competitive ELISA methods. In addition, possible biotin interactions were investigated by determining the concentrations of all the study parameters in serum pools containing different amounts of biotin.
Results
More consistent results were obtained with biotin-competitive, ghrelin-competitive, KISS1-competitive, GHSR, leptin and LEPR ELISA kits. Unexpectedly, high levels were detected in the biotin sandwich ELISA kit, indicating that biotin interference may also occur in manually run research kits.
Conclusions
Biotin exhibited an interference effect even in well-functioning, qualified kits, and this negative effect was less common in competitive kits. Biotin interference was closely associated with the quality of the research kit, the parameters studied and the presence of high biotin concentrations in the blood.