Liquid Biopsy

Multicentre verification does not define every negative sample

by Tala Benor

Standardized analytical verification across dPCR and NGS platforms addresses only part of ctDNA negativity. For each site and platform, negative calls still need the tested DNA input, reportable limit at that input, variant coverage, and specimen handling window. In longitudinal testing, baseline ESR1 detectability and collection time relative to therapy determine whether absence can support molecular change rather than nonevaluability.

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Signal / Noise

The abstract gives concrete limits behind that distinction: dPCR used 10 to 30 ng DNA per reaction, with detection limits of 0.01% to 0.07% variant allele frequency; NGS limits ranged from 0.01% to 0.1%, depending on platform and input. Those ranges describe analytical verification with reference materials, focused on p.Y537S and p.D538G. They therefore provide conditions for comparing assays, but the abstract doesn't establish when a patient's longitudinal negative result is evaluable. [Study abstract](https://pubmed.ncbi.nlm.nih.gov/42307899/)

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