Molecular Diagnostic Assay Validation - Update to the 2009 AMP Molecular Diagnostic Assay Validation White Paper

Sep 30, 2026

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Provides a practical reference for validating molecular diagnostic assays for clinical laboratory implementation, updating the 2009 AMP Molecular Diagnostic Assay Validation White Paper. Outlines goals and stepwise documentation expectations, contrasts assay validation and assay verification under CLIA and CAP, and specifies required performance characteristics for unmodified FDA-approved/cleared tests versus modified or non-FDA-cleared tests. Includes guidance for assay design and clinical validation, emphasizing clinical validity and utility. Mentions a sample summary checklist template for laboratory director review and sign-off.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages11

Reading Time28 min

AccessFree to download

Introduction

Summary

Definitions

Assay Validation

Assay Verification

Assay Types

Unmodified FDA-Approved or Cleared Tests

Modified FDA-Approved or non-FDA Cleared Tests

Assay Design

Clinical Validation

Molecular Diagnostic Assay Validation Update to the 2009 AMP Molecular Diagnostic Assay Validation White Paper Members of the 2013 and 2014 Clinical Practice Committees of the Association for Molecular Pathology The 2013 AMP Clinical Practice Committee consisted of Matthew J. Bankowski, Milena Cankovic, Jennifer Dunlap, Larissa V. Furtado*, Jerald Gong, Thomas Huard, Linda Jeng*, Loren Joseph (Chair), Annette Kim, Bryan Krock, Marilyn M. Li, Mary Lowery-Nordberg, Melissa Miller, Caroline Sue Richards, and Paul Rothberg. * Project leads Standard of practice is not defined by this article, and there may be alternatives. See Disclaimer for further details. INTRODUCTION The goal of method validation in the molecular diagnostics laboratory is to ensure that a given test is ready for implementation in the clinical laboratory. To reach that goal, each step of the testing process must be carefully evaluated and documented. Such validation is relatively standardized for high volume automated assays in fields such as clinical chemistry. It is challenging to apply those standardized practices to molecular diagnostics laboratories, which rely heavily on low-volume labor-intensive tests, both FDA approved and laboratorydeveloped. Excellent and comprehensive documents have recently been published on this topic 1-10 . This overview is intended to provide a brief practical reference that can be used to guide the validation process in molecular diagnostics laboratories. In addition, a sample summary checklist is provided in the appendix as a template to facilitate documentation of laboratory director review and sign off on the validation process when new assays are developed or adopted. SUMMARY CLIA (42 CFR 493 . 1253) and CAP (MOL 30785-31705) require that laboratories validate the performance of tests before reporting patient results. DEFINITIONS Assay Validation • WHO definition: “the action (or process) of proving that a procedure, process, system equipment or method used works as expected and achieves the intended result” 11. • FDA definition: “confirmation by examination and provision of objective evidence that the particular requirements for a specific intended use can be consistently fulfilled”12. • CLSI definition: “confirmation by examination and provision of objective evidence through a defined process that the particular requirements for a specific intended use can be consistently fulfilled.”13 This definition is also used by CAP Accreditation Programs 14 . Assay Verification • CLIA definition: “confirmation by examination and provision of objective evidence that specified requirements have been fulfilled”13, or “confirmation that the laboratory using a test can replicate the manufacturer’s performance claims when the test is used according to the package insert” 15. • CAP definition: “an abbreviated process to demonstrate that a test performs in substantial compliance to previously established claims”14 Verification applies to unmodified non-waived (moderate-and high-complexity) tests that have been cleared or approved by the FDA and are labeled “for in vitro diagnostic use.” ASSAY TYPES UNMODIFIED FDA-Approved or Cleared Tests – Requirements For UNMODIFIED FDA-approved or FDA-cleared tests, laboratories must verify that test(s) perform(s) as expected by obtaining data on: • Accuracy • Precision • Reportable range • Linear range (for quantitative assays) • Reference intervals (normal values) for laboratory patient population These performance characteristics are published in the manufacturer’s package insert. MODIFIED FDA-Approved or non-FDA Cleared Tests – Requirements For MODIFIED FDA-Approved tests or for non-FDA cleared tests (e.g., Laboratory Developed Procedures (LDP), previously LDTs) laboratories must establish the following performance characteristics: • Accuracy • Precision • Reportable range • Linear range (for quantitative assays) • Reference intervals

FAQs

What is the primary goal of method validation in a molecular diagnostics laboratory?

To ensure a given test is ready for clinical laboratory implementation by carefully evaluating and documenting each testing step.

How do assay validation and assay verification differ?

Assay validation focuses on proving a method works as expected for intended use and achieving specified requirements consistently, while assay verification confirms objective evidence that specified requirements are fulfilled and that the lab can replicate manufacturer performance claims for the intended use.

What performance characteristics are required for unmodified FDA-approved or cleared tests?

Laboratories must verify accuracy, precision, reportable range, linear range for quantitative assays, and reference intervals for the laboratory patient population.

Sep 30, 2026

Upload by Kyle