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Detecting KRAS mutations in pancreatic cystic neoplasms: droplet digital PCR versus targeted next-generation sequencing

Published:October 15, 2022DOI:https://doi.org/10.1016/j.hpb.2022.10.007
      DNA-based testing of pancreatic cyst fluid (PCF), obtained by endoscopic ultrasound fine needle aspiration (EUS-FNA) may improve diagnostic accuracy for differentiation between types of pancreatic cystic neoplasms (PCN),
      • Wu J.
      • Jiao Y.
      • Dal M.M.
      • Maitra A.
      • de Wilde R.F.
      • Wood L.D.
      • et al.
      Whole-exome sequencing of neoplastic cysts of the pancreas reveals recurrent mutations in components of ubiquitin-dependent pathways.
      which is currently suboptimal. Mutations in DNA (a.o. KRAS, GNAS) can differentiate non-mucinous from (pre)-malignant, mucinous PCN. Various techniques are available to detect mutations in DNA; including targeted next generation sequencing (NGS), like Ion Torrent NGS, and since more recently droplet digital PCR (ddPCR). Molecular tests involving ddPCR are not widely used in clinical setting. DdPCR requires only 1 ng of DNA for a 0.1% sensitivity,
      • Demuth C.
      • Spindler K.G.
      • Johansen J.S.
      • Pallisgaard N.
      • Nielsen D.
      • Hogdall E.
      • et al.
      Measuring KRAS mutations in circulating tumor DNA by droplet digital PCR and next-generation sequencing.
      and is therefore a potentially suitable technique for DNA-based testing of PCF, which tends to contain low DNA concentrations. In comparison, NGS is able to detect mutations in multiple genes simultaneously
      • Singhi A.D.
      • Nikiforova M.N.
      • McGrath K.
      DNA testing of pancreatic cyst fluid: is it ready for prime time?.
      on around 10 ng of DNA and has a limit of detection of 3–5%.
      • Stasik S.
      • Schuster C.
      • Ortlepp C.
      • Platzbecker U.
      • Bornhäuser M.
      • Schetelig J.
      • et al.
      An optimized targeted Next-Generation Sequencing approach for sensitive detection of single nucleotide variants.
      The aim of this pilot study was to evaluate whether ddPCR is suitable for measuring KRAS-mutations in PCF, and to compare it with NGS.
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