Heidi Hyytiä1, Taina Heikkilä2, Pirjo Hedberg3, Tarja Puolakanaho3, Kim Pettersson2. 1. Department of Biotechnology, University of Turku, Tykistökatu 6 A, Turku 20520, Finland. Electronic address: heidi.hyytia@utu.fi. 2. Department of Biotechnology, University of Turku, Tykistökatu 6 A, Turku 20520, Finland. 3. NordLab Oulu, Oulu University Hospital, Oulu, Finland; Department of Clinical Chemistry, University of Oulu, Oulu, Finland.
Abstract
OBJECTIVES: To study the skeletal troponin I (skTnI) cross-reactivity of four different commercially available antibodies in four cardiac troponin I (cTnI) research assay versions having the same epitope specificity as evidenced by peptide mapping. DESIGN AND METHODS: The four research assays all use two solid phase antibodies and one detection antibody attached to intrinsically fluorescent nanoparticles. Two alternative antibodies were used for one capture antibody and two for the detector antibody. The assays were evaluated in terms of analytical sensitivity and by determining assay cross-reactivity to skTnI. Additionally, regression analysis was performed by measuring a sample panel (n=101) with all of the four assay versions. RESULTS: A false-positive cTnI concentration of >7000ng/L was measured with one of the assay versions, when serum was spiked with 500,000ng/L skTnI. The corresponding observed cTnI values for the other three assay versions varied from 616ng/L to 727ng/L. Out of the 101 clinical samples assayed, five showed spuriously (3- to 148-fold) elevated cTnI values with the skTnI interference prone assay setup, but not with the other assay versions. According to our investigational skTnI assay, all five samples contained measurable amounts of skTnI (range: 5500-702,000ng/L). CONCLUSIONS: Two out of four cTnI antibodies tested cross-reacted vastly with skTnI but did not cause any notable interference unless paired together. Therefore, skTnI cross-reactivity should be carefully assessed when cTnI assay antibodies claimed to be cTnI specific are selected.
OBJECTIVES: To study the skeletal troponin I (skTnI) cross-reactivity of four different commercially available antibodies in four cardiac troponin I (cTnI) research assay versions having the same epitope specificity as evidenced by peptide mapping. DESIGN AND METHODS: The four research assays all use two solid phase antibodies and one detection antibody attached to intrinsically fluorescent nanoparticles. Two alternative antibodies were used for one capture antibody and two for the detector antibody. The assays were evaluated in terms of analytical sensitivity and by determining assay cross-reactivity to skTnI. Additionally, regression analysis was performed by measuring a sample panel (n=101) with all of the four assay versions. RESULTS: A false-positive cTnI concentration of >7000ng/L was measured with one of the assay versions, when serum was spiked with 500,000ng/L skTnI. The corresponding observed cTnI values for the other three assay versions varied from 616ng/L to 727ng/L. Out of the 101 clinical samples assayed, five showed spuriously (3- to 148-fold) elevated cTnI values with the skTnI interference prone assay setup, but not with the other assay versions. According to our investigational skTnI assay, all five samples contained measurable amounts of skTnI (range: 5500-702,000ng/L). CONCLUSIONS: Two out of four cTnI antibodies tested cross-reacted vastly with skTnI but did not cause any notable interference unless paired together. Therefore, skTnI cross-reactivity should be carefully assessed when cTnI assay antibodies claimed to be cTnI specific are selected.
Authors: Vincent L Aengevaeren; Aaron L Baggish; Eugene H Chung; Keith George; Øyunn Kleiven; Alma M A Mingels; Stein Ørn; Rob E Shave; Paul D Thompson; Thijs M H Eijsvogels Journal: Circulation Date: 2021-12-13 Impact factor: 29.690