Abstract
Background
In vitro specific IgE (sIgE) testing has become an important tool for the diagnosis of IgE-mediated allergic diseases. Current methods used to detect allergen sIgE are time consuming and/or expensive. Therefore, a new method was developed for rapid quantitative detection of cat dander-sIgE antibody based on homogeneous chemiluminescence immunoassay.
Methods
Selection of chemibeads with different chemical groups, and the best Light-initiated chemiluminescence assay (LiCA) analytical mode for cat dander-sIgE detection. To validate and eliminate the interference of IgE on the detection of cat dander-sIgE, concentration of biotinylated anti-human IgE antibody was optimized. For quantification of cat dander-sIgE, a calibration curve was established, and the performance of the assay was evaluated according to clinical guidelines.
Results
Indirect LiCA is the best mode of analysis and biotinylated anti-human IgE antibody at a dilution ratio of 1:250 minimizes IgE interference. The coefficient of variation of the developed LiCA was 1.49% to 4.66%, with an intermediate precision of 6.90% to 8.21%. The LoB, LoD, and LoQ of the assay were 0.023 kUA/L, 0.056 kUA/L and 0.185 kUA/L. The coefficient of correlation (r) between LiCA and ImmounoCAP was 0.9478.
Conclusions
A cat dander-sIgE quantitation assay based on homogeneous chemiluminescence immunoassay was established, which could be a new reliable analytical tool for the determination of cat dander-sIgE.
Acknowledgments
We thank Clinical Laboratory of Tianjin Children’s Hospital and Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine for material support.
Author contributions
All authors have accepted responsibility for the entire content of this submitted manuscript and approved the submission. Yuanmin Sun designed the study, analyzed the data, and wrote the manuscript. Ren Na and Lina Zhu analyzed the data and wrote the manuscript. Xiaohui Yang and Xin Tan performed the assays and helped analyze the data. Yang Yu and Huiqiang Li helped analyze the data and review the manuscript.
Declaration of interest
The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.