High agreement between one-stage clot-based and chromogenic factor VIII activity assays

INTRODUCTION: A number of gene therapy studies have shown discrepancies between the one-stage and chromogenic factor VIII (FVIII) assays. Studies comparing the two assays are of small sample size. We aimed to compare the one-stage clot-based assay with the chromogenic assay in measuring FVIII activity in blood sample in a single tertiary academic centre.

METHODS: This is a single-centre retrospective study conducted at Sultan Qaboos University Hospital. Both assays performed FVIII measurements at the SQUH from March 2009 to March 2018. A total of 3623 samples from a range of haematologic conditions had both assays performed.

RESULTS: The median age of all patients was 26 years (IQR: 17-37). The mean FVIII activity by one-stage clot-based and chromogenic assays were 1.2 IU/ml (SD: 0.6) and 1.2 IU/ml (SD: 0.5), respectively. Bland-Altman analysis showed excellent agreement with a very minimal bias of -0.0034. High baseline FVIII activity and the diagnosis predicted disagreement between the two assays ( P values < 0.001). Positive ANA and FVIII inhibitor had no impact on the disagreement ( P value = 0.2 and 0.7, respectively). A subgroup analysis of samples with FVIII activity of <0.4 IU/ml showed 21.1% discrepancy with the majority (75.8%) of samples showing overestimated results by the OSA.

CONCLUSION: One-stage clot-based and chromogenic FVIII assays showed excellent agreement for measuring FVIII activity.