Establishment of a new protein C detection system based on chromogenic substrate assay and its clinical diagnostic value for deep vein thrombosis.

Abstract

BackgroundDeficiency of protein C (PC) affects the balance between blood coagulation and fibrinolysis in the human body. Chromogenic‐based assay is recommended as the preferred screening method for detecting PC deficiency. We established a PC detection system based on the chromogenic substrate assay.MethodsFirst, a kit for the determination of PC activity in plasma was elaborately developed and its reaction parameters on XL‐3200c were explored. Then, we evaluated its performance and collected specimens to compare the test results obtained with those of the Siemens detection system. Finally, the clinical diagnostic efficacy of this detection system for deep vein thrombosis (DVT) was assessed.ResultsOptimum conditions for PC detection were 0.25–0.1 U/ml protein C activator Protac® and 2.5–1 mM Pefachrome®PCa5297. The composition and concentration ranges of buffer substances and stabilizers in the kit were also explored. Satisfactory results were observed in performance evaluation. The test results of the newly built detection system were highly correlated with those of the Siemens detection system (R 2 = 0.9771 in the control group and R 2 = 0.9776 in the DVT group), and Bland‐Altman plots also showed high consistency between the two detection systems. In addition, the area under the curve (AUC) of the newly built PC detection system for DVT was 0.888, indicating this system could effectively improve the diagnostic sensitivity and specificity for DVT.ConclusionIn this study, a sensitive, wide linear range and reliable PC activity detection system were established.