A novel combination mode for inhalant allergen screening based on 8-well microplates by LiCA

Currently, allergen detection is primarily classified into two types: single detection and multiple combined detection. The challenges include large volume of serum required, high testing costs, and fixed combinations of test items. A cost-effective method that uses a small amount of serum and allows for the flexible combination of allergen screening is necessary. Light-initiated Chemiluminescence Assay (LiCA) requires no washing, only need little serum, and supports flexible combination detection of allergens.

Based on the advantages of LiCA technology, which does not require washing, the '4 + 3X + 1' allergen detection mode was established in this study. The antigens are coated on chemibeads, which are selectively added into the detection wells to achieve flexible combination, supporting the simultaneous detection of eight allergens.

This study introduces a new allergen screening model termed "4 + 3X + 1", where "1" represents tIgE, "4" comprises four commonly encountered indoor inhalable allergens, and "3X" encompasses a variety of frequent inhalable allergens. Allergens from "3X" could be selected based on regional characteristics and patient-specific conditions. We tested the intermediate precision(3.59 %-9.71 %) and repeatability(2.81 %-9.31 %) of the four allergens in "4-fixed", and LoB(0.035-0.066 kU_A/L), LoD(0.092-0.156 kU_A/L), and LoQ(0.135-0.199 kU_A/L) all showed good results. We compared the consistency of LiCA and ImmunoCAP/Dot-ELISA, each of which was greater than 0.8828. In addition, two allergens were selected from "4-fixed" and "3X-optional "respectively to detect the sensitivity(80 %-97.72 %) and specificity(93.75 %-100 %).

The proposed combined test model offers a new, economical, and rapid option for allergen screening that requires low blood volumes and can be customized for individual patients.