AOAC Guidance on FA Immunoassay Validation (November 2023)

514 Figure 3: Design 2a. Lot: test kit lot, TP: test portion, E: ELISA (well) measurement. Design 2a can be 515 used to estimate intermediate precision, repeatability, and ELISA variance.

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5.2.4.2.3.8. Design 2b

5.2.4.2.3.8.1. Design 2b ( Figure 4 ) can be used to estimate (1) intermediate precision (which includes repeatability, test kit lot variance (with 2 df), day/operator confounded variance, and ELISA variance), repeatability (which includes test portion variance and ELISA variance), (3) ELISA 5.2.4.2.3.8.2. Three test kit lots are used to analyze each test material. Two operators conduct analysis on two days for each test kit lot. For each day and lot, the assigned operator conducts extraction and analysis of two test portions of test material, with two ELISA measurements (i.e., wells) performed per test portion. For each test portion, the entire analysis variance, and (4) lot-to-lot product consistency.

procedure is conducted within the corresponding day.

530 Figure 4: Design 2b. Lot: test kit lot, TP: test portion, E: ELISA (well) measurement. Design 2b can be 531 used to estimate intermediate precision, repeatability, ELISA variance, and lot-to-lot product 532 consistency. 533