Comparison of clot-based and chromogenic assay for the determination of protein c activity

Activated protein C inactivates factor Va and VIIIa. Deficiency of this natural anticoagulant may result in recurrent venous thrombosis. Performance characteristics of clot-based and chromogenic protein C activity assays are different. The clot-based assay has limitations because of interference with coagulation inhibitors resulting in spuriously increased protein C levels or underestimation because of elevated levels of factor VIII and Factor V-Leiden mutation. The chromogenic assay is not influenced by such interferences but only detects functional defects of protein C that involve the active site rendering it insensitive to rare mutations. To compare two methods, we conducted a retrospective study from January 2015 to June 2017. Our results showed a good correlation between clot-based and chromogenic assay (R = 0.94 and r(2) = 0.88). The study of agreement between the two methods by the Bland–Altman method showed that chromogenic method on an average measures 7.8% more protein C than that of clot-based. The results also showed that the bias between the two methods is significant. The positive trend noted was contributed by the values of less than 20% of protein C. Both clot-based and chromogenic assays had high sensitivity; however, the chromogenic assay showed better specificity (97%) as compared with the clot-based assay (93%). In conclusion, we recommend the chromogenic method as the assay of choice, which is also recommended by the College of American Pathologist Consensus Study over activated partial thromboplastin time-based assay. We have shown here that despite a good correlation between the two techniques, there is a difference as highlighted by the difference plots.