Washing-free Chemiluminescence Immunoassay for Rapid Detection of cTnI in Whole Blood Samples

Abstract

Background: Chemiluminescence immunoassay (CLIA) has always been a great challenge in detecting whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Results: In this scheme, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotin-conjugated cTnI antibody and detected by streptavidin/acridine aster-conjugated PCMS. After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the cTnI concentrations of the serum samples, plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at 0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62~5.67%. The assay was linear over the studied range of 0.01–50.00 ng/mL, and no hook effect was found when cTnI concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit (Abbott assay kit), the correlation coefficient was 0.9859.Conclusions: A washing-free chemiluminescence immunoassay (CLIA) was established for the rapid detection of cardiac troponin I (cTnI) in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and polychloromethylstyrene microspheres (PCMS) for signal amplification, which showed great potential in clinical application.