AFP-CHM-ELISA

We have thus developed AFP-CHM-ELISA — a highly sensitive ELISA for AFP detection, by immobilizing a chitosan hydrogel membrane to polystyrene. The performance of the method was further evaluated using clinical serum samples.
The linear range, limit of detection (LOD), inter-batch and intra-batch precision, recovery tests, and other parameters were determined for the newly developed method, while its performance metrics such as sensitivity, repeatability and accuracy were evaluated according to the standardized validation protocol, which provides validated procedures for evaluating analytical method characteristics (Chan et al., 2004). Furthermore, an accelerated stability test and real-time stability test were conducted based on the CLSI EP25-A [28].
The AFP-CHM-ELISA was used to detect clinical samples, and its performance was compared with traditional ELISA and chemiluminescence methods, followed by linear regression and reliability analyses.
The OD value of calibrator A was assigned as the negative control (N), while calibrators B to F were designated as positive controls (P). Sensitivity was calculated as the ratio of the OD value of calibrator B to that of calibrator A (PB/N), and the linear range was determined by the ratio of the OD value of calibrator F to that of calibrator A (PF/N).
The correlation between the two methods was assessed using the regression equation and correlation coefficients. Method agreement was evaluated using Bland-Altman analysis.
All statistical analyses were performed using GraphPad Prism (Version 9.0, USA), with a significance level of p < 0.05.