AN INTERNATIONAL INTER-LABORATORY DIGITAL PCR STUDY DEMONSTRATES HIGH REPRODUCIBILITY FOR THE MEASUREMENT OF A RARE SEQUENCE VARIANT
Alexandra S Whale,
Alison S Devonshire,
Gerwyn M Jones,
Andreas W Berger,
Nina Dahl Kjersgaard,
Rikke Fredslund Andersen,
Álvaro González Hernández,
Charles A. Haynes,
Carole A Foy,
Jim F. Huggett
Posted 28 Sep 2016
bioRxiv DOI: 10.1101/077917 (published DOI: 10.1021/acs.analchem.6b03980)
Posted 28 Sep 2016
This study tested the claim that digital PCR (dPCR) can offer highly reproducible quantitative measurements in disparate labs. Twenty-one laboratories measured four blinded samples containing different quantities of a KRAS fragment encoding G12D, an important genetic marker for guiding therapy of certain cancers. This marker is challenging to quantify reproducibly using qPCR or NGS due to the presence of competing wild type sequences and the need for calibration. Using dPCR, eighteen laboratories were able to quantify the G12D marker within 12% of each other in all samples. Three laboratories appeared to measure consistently outlying results; however, proper application of a follow-up analysis recommendation rectified their data. Our findings show that dPCR has demonstrable reproducibility across a large number of laboratories without calibration and could enable the reproducible application of molecular stratification to guide therapy, and potentially for molecular diagnostics.
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