Fully automated PCR detection of KRAS mutations on pancreatic endoscopic ultrasound fine-needle aspirates

Abstract

This is the first study that applied the novel Idylla KRAS test to the clinical setting of pancreatic cancer. In particular, this system can be easily implemented in the routine assessment of pancreatic EUS-fine-needle aspiration-derived DNA samples to quickly provide information on KRAS mutational status to supplement cytological evaluation.

Evaluation of a fast and fully automated platform to diagnoseEGFRandKRASmutations in formalin-fixed and paraffin-embedded non-small cell lung cancer samples in less than one day

“…Nevertheless, rare EGFR and KRAS variants and mutations in other molecular targets are not detected by this fast diagnosis platform to date. In previous studies, Idylla did not detect double mutants (eg, KRASG12V and KRASQ61H mutant) but only reported the mutation with the smallest ΔCq per assay/cartridge 16. In our study, Idylla identified every KRAS and EGFR activating mutations including two samples with a double mutation ( EGFRG719X and EGFRS768I mutations in case #1; EGFRG719X and EGFRL861Q in case #9).…”

Idylla assay and next generation sequencing: an integrated EGFR mutational testing algorithm

“…Our data showed that, when clinical decisions should be taken quickly and when saving time is a crucial issue, since days have been spent for an ‘invalid’ NGS result, Idylla can provide a solution. As a general rule, regardless of the sample type, we directly pipetted in the cartridge from 3 up to 10 µL of extracted DNA preparation 4. Indeed, in this study, Idylla ‘rescued’ a large portion of cases (20/25; 80%) whose preliminary assessment by NGS was unfeasible (figure 4).…”

“…Although this procedure had been designed by the manufacturer to process 5–10 µm FFPE tissue sections, we have previously shown that archival extracted DNA can directly be pipetted inside an EGFR Idylla cartridge 4. Cases with valid NGS results were tested by pipetting 3 µL (corresponding to 1/10th of the original DNA preparation from the sample).…”

EGFRmutation detection on lung cancer cytological specimens by the novel fully automated PCR-based IdyllaEGFRMutation Assay

“…The instrument is composed of a sample preparation module integrated with a combined PCR thermocycling and fluorescence detection module. All consumables required to perform sample preparation and RT-PCR detection are provided in a single disposable cartridge that is loaded into the Idylla system to enable the simultaneous detection of up to 30 molecular targets from a variety of solid and liquid sample types, including DNA preparations from routine cytological specimens 10. Closing of the cartridge after inserting the sample avoids cross-contamination.…”

“…More recently, Biocartis developed an Idylla EGFR test prototype. Although, the Idylla tests were designed for use with formalin-fixed paraffin-embedded (FFPE) sections, the Idylla system can also process DNA preparations from cytological samples, as previously shown 10. The aim of the present study was to assess the Idylla EGFR test performance on lung cancer cytological specimens.…”