B-058 Development and Evaluation of the Assay for Detection of Human Specific IgE Against Animal Dander Allergens on a Fully Automated Chemiluminescence Immunoassay Platform—Autolumo® A2000 Plus

Abstract

Abstract Background The detection of specific IgE is a key method for diagnosing allergic diseases. An accurate, user friendly and cost-effective way to clinically diagnosing allergies is beneficial for the patients. Our detection kit together with the Autolumo A2000 Plus chemiluminescence fully automated platform provide a new and more effective (40 minutes) way for detection of specific IgE with simple operation, high sensitivity and specificity. Methods The Animal Dander Allergen Mix Assay kit detects specific IgE against cat dander (E1), horse dander (E3), cow dander (E4) and dog dander (E5). Performance evaluation of the animal dander mix, E1, E3, E4 and E5 assay kits for the detection of specific IgE in serum was performed on our automated chemiluminescence platform (Autolumo A2000 Plus) and compared to a well-characterized platform (ImmunoCAP, Phadia). During detection, magnetic particle suspension, antigen solution, patient sample, and enzyme conjugate were added sequentially. A streptavidin-biotinylated antigen-sIgE antibody-anti-IgE-HRP conjugate complex is formed, wherein the amount of the anti-IgE-HRP conjugate is positively correlated to the amount of sIgE antibodies in the patient sample. After the substrate is added, the HRP catalyze the chemical reaction to generate a recordable signal in the form of light. The intensities of the light were obtained by using a chemiluminescence quantitative analyzer, and the standard curve was generated according to the 7-point calibration, and then the sIgE antibody concentrations in the serum sample were analyzed. The overall agreement, method comparison, precision, linearity, and stability of our mixed and single assay kits were evaluated using serum samples (n > 140) that had been characterized with Phadia. Results The overall agreement for the animal dander mix assay is 91.84%, with the average intra-run and inter-run precision to be 1.97% and 4.21% respectively. The limit of blank (LoB) and limit of detection (LoD) for the animal dander mix assay is 0.019 kU/L and 0.045 kU/L respectively. The sensitivity for our single E1, E3, E4, and E5 assays are 97%, 85.6%, 70.8%, and 80.2%, respectively. All single assays have a specificity >90%, total precision <5% CV, and a linearity with R2 > 0.999. The LoB, LoD and recommended LoQ for all single assays are 0.019 kU/L, 0.037 kU/L, and 0.1 kU/L respectively. The average signal drifts after 10 days incubation in 37C is less than 10% indicating the assays are stable. Conclusion Autolumo A2000 Plus is a fully automated chemiluminescence Immunoassay platform that provides accurate and cost-effective screening for allergy diagnostics with excellent overall agreement, precision, and stability.