Analytical Validation of qPCR-Based Multivariate Index Assays in a Clinical Laboratory: Practical Challenges and Limitations

摘要: Background: Multivariate index assays (MIAs) to evaluate disease status and/or therapeutic efficacy are increasingly being used in clinical laboratories as laboratory-developed tests (LDTs). Before use, diagnostic and analytical performance specifications of LDTs must be established. Several regulatory guidelines have been published that address specific components validation procedures, but the interpretation for MIAs is ambiguous creates confusion when implementing a novel MIA laboratory. Content: CLSI methods were evaluated develop strategy establish sensitivity, precision, specificity, stability qPCR-based MIAs. Limitations challenges identified while evaluating literature this discussed review, including sample sourcing integrity, laboratory contamination, throughput. Due diversity MIAs, we discuss additional considerations researchers intending transfer Summary: A practical assess characteristics was developed tested before use. important limitations, challenges, during development procedures not addressed or literature. The described can provide insight future developers LDTs.