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Establishment of a microsphere-based flow cytometric assay for quantitative detection of total immunoglobulin E
Qi Jiang1, Yuanmin Sun2, Xiaoming Cui3
1Department of Basic Medical Sciences, Tianjin Medical College, Tianjin 300222, China.
Background:
In the diagnosis of allergic diseases, in vitro serum IgE level tests were conventionally used. In recent years, microsphere-based flow cytometric assays for IgE have gained more attention. The present study aimed to establish a microsphere-based flow cytometric assay for quantitative detection of tIgE.
Methods:
The assay was established after selection of microsphere, antibody pair, detection mode, and optimization of coating ratio. In order to quantitate tIgE, the calibration curve was established. The detection performance of this new tIgE flow cytometric assay was also confirmed. In addition, the correlations between this assay with the ImmunoCAP assay or the immunoturbidimetry assay were evaluated.
Results:
This new tIgE flow cytometric assay has low CVs of repeatability (7.62 % and 6.98 %) and intermediate precision (7.50 % and 8.41 %). The detection range was 2 ∼ 6000 IU/mL. The range of linearity was from 2 ∼ 3363 IU/L (r = 0.9932). The correlation coefficient (r) for the correlation analysis between the results of the newly developed tIgE flow cytometric assay and ImmunoCAP assay or immunoturbidimetry assay was 0.9345 and 0.9348, respectively.
Conclusion:
A tIgE flow cytometric assay was successfully established. It could be used in clinical laboratories and quantitate lots of allergen-sIgE detected by flow cytometric assay.
