This document outlines general requirements for the validation and verification of multiplex molecular tests that detect multiple nucleic acid target sequences simultaneously. Applicable to both qualitative and quantitative in vitro diagnostic (IVD) methods and laboratory-developed tests (LDTs), it provides essential guidance for detecting human or microbial nucleic acid targets in clinical specimens. It supports medical laboratories, IVD developers, biobanks, research institutions, and regulatory authorities in ensuring reliable molecular testing. This document does not cover metagenomics. Laboratory-developed or in-house tests refer to examination procedures developed for a laboratory’s own use.