A Novel Rapid Single-Integrated Immunoassay for Plasma Thrombomodulin: Analytical Performance and Multi-Center Clinical Validation

Abstract

Thrombotic diseases have emerged as a significant public health issue, with timely detection, diagnosis, and treatment being closely associated with patient prognosis. Consequently, the identification of highly sensitive and specific biochemical markers for thrombotic diseases holds substantial clinical importance. In this study, we developed a rapid, single-integrated quantitative detection method for thrombomodulin (TM). This method integrates all reagent components onto a single, sophisticated card strip, enabling the completion of the assay in merely 20 minutes. Such efficiency enhances the potential for early screening of thrombosis within community and primary care medical institutions. The analytical performance results indicate that the limit of blank (LoB), limit of detection (LoD), and limit of quantitation (LoQ) were determined to be 0.5, 0.8, and 1.2 TU•mL -1 , respectively. The repeatability and precision of the method were found to be significantly below 10%, with a linear range extending from 1.2 to 200 TU•mL -1 . Accuracy evaluations revealed relative biases of -3.35% and 4.79% for the two reference materials tested. Furthermore, clinical trials conducted across two clinical research centers involved a total of 236 clinical plasma samples. The results of the clinical analysis demonstrated excellent consistency with the reference reagent (Sysmex). The receiver operating characteristic (ROC) curve and significance analysis indicated that this method possesses commendable specificity and sensitivity. In summary, the single-integrated immunoassay method developed in this study represents a rapid, convenient, accurate, and sensitive detection approach. It is particularly well-suited for the early screening and diagnosis of thrombotic diseases in community and primary healthcare institutions.