Isotype-specific agglutination-PCR (ISAP): A sensitive and multiplex method for measuring allergen-specific IgE

• Published in: Journal of allergy and clinical immunology Vol. 141; no. 5; pp. 1901 - 1904.e15
• Main Authors: Tsai, Cheng-ting, Mukai, Kaori, Robinson, Peter V., Gray, Melissa A., Waschmann, Malika B., Lyu, Shu-Chen, Tsai, Mindy, Chinthrajah, Rebecca S., Nadeau, Kari C., Bertozzi, Carolyn R., Galli, Stephen J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2018; Elsevier Limited
• Subjects: Agglutination; Agglutination - immunology; Allergens; Allergens - immunology; Allergies; Anaphylaxis; Animals; Antibody Formation - immunology; Deoxyribonucleic acid; DNA; Female; Hypersensitivity - immunology; Immunoglobulin E; Immunoglobulin E - immunology; Immunoglobulins; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Mice, Inbred C57BL; Polymerase Chain Reaction - methods
• ISSN: 0091-6749; 1097-6825; 1097-6825
• DOI: 10.1016/j.jaci.2017.11.021

[...]we used CLIQ to analyze plasma obtained from 20 peanut-allergic subjects upon their enrollment into the institutional review board–approved peanut oral immunotherapy study: safety, efficacy and discovery trial (ClinicalTrials.gov Identifier: NCT02103270).9 CLIQ results displayed substantial positive correlations with ImmunoCAP data from the same specimens (Fig 2, E; see Figs E9, E10, and E11 in this article's Online Repository at www.jacionline.org). DNA sequences and primers were optimized to minimize the formation of secondary structures and primer dimers while maximizing amplification efficiency.Analysis of human plasma Plasma samples from patients with peanut allergy were obtained as part of their enrollment into an institutional review board–approved clinical trial of oral immunotherapy in children and adults with peanut allergy (peanut oral immunotherapy study: safety, efficacy and discovery; ClinicalTrials.gov Identifier: NCT02103270). [...]we calculated LODs of 0.95, 0.55, and 1.94 (ΔCt units) for anti–Ara h 1, anti–Ara h 2, and anti–Ara h 3 sIgEs, respectively (indicated by the vertical red dashed lines in Fig E9). The number of antibody molecules in each measurement was calculated as (1 × 10−6 L) × Antibody concentration (M). [...]the number of antibodies in each dilution series ranged from 10−13 to 10−18 mol.