Table 1

Details of the available BRAF, KRAS and NRAS assays on the Idylla system compared with common PCR (Cobas) and NGS (Ion Torrent) platforms

Gene targetIdylla14Cobas z 480 (PCR)32Ion PGM (NGS)23
BRAF1) NRAS-BRAF-EGFRS492R Mutation Assay20
Coverage: V600D/E/E2/K/R
PCR method: PlexPrime/PlexZyme Cq curve
Detection limit: ≤5%
Turnaround time: 2 hours (approx.)
2) BRAF Mutation Test19
Coverage: V600D/E/E2/K/M/R
PCR method: TaqMan
Detection limit: 1%
Turnaround time: 1.5 hours
1) BRAF V600 Mutation Test26
Coverage: V600E
PCR method: TaqMan Cq curve
Detection limit: >5%
Turnaround time: <8 hours
AmpliSeq Cancer Hotspot Panel v224
Coverage*: R444W, P453T, R462I, G464E/V/R, G466R/V/A/E, G469R/S/V/A/E, V471F, N581S, I582M, F583F, L584L/F, E586E/K, D587E/A, I592M/V, D594E/N/V/G, F595S/L, G596D/R, L597L/Q/S/V/R, A598V, A598_T599insV, T599I, T599_V600insTT, T599_V600insT, V600D/E/G/A/K/R/M/L/Q, K601N/E, V600_K601>E, K601del, R603†, W604G, W604del S605F/N/G, G606E, H608R
Detection limit: 98% detection rate for 5% variant frequency at positions with average sequencing coverage from 1000X to 4000X
Turnaround time: single day
KRASKRAS Mutations Test18
Coverage: G12C/R/S/A/D/V, G13D, A59E/G/T, Q61H/H2/K/K2/L/R, K117N1/N2, A146P/T/V
PCR method: PlexPrime/PlexZyme Cq curve
Detection limit: ≤5%
Turnaround time: 2 hours (approx.)
KRAS Mutation Test33
Coverage: G12C/R/S/A/D/V, G13C/S/R/D/V/A, Q61K/E/P/R/L/H/H2
PCR method: TaqMelt melting curve
Detection limit: <5%
Turnaround time: <8 hours
AmpliSeq Cancer Hotspot Panel v224
Coverage*: V8V, G10_A11insG, G12_G13insG, A11P/V, A11_G12insGA, G12F/C/L/V/S/R/E/V/D/A/G/Y/I/W, G13R/C/S/R/V/D/A/V/G/E, G13_V14insG, V14I, G15S, A18D, L19F, Q22K, T58I ,A59G/T/E, Q61K/E/P/R/L/H/K, K117N, A146T/V/P
Detection limit: see above
Turnaround time: single day
NRASNRAS-BRAF-EGFRS492R Mutation Assay20
Coverage: G12C/S/D/A/V, G13D/V/R, A59T, Q61K/L/R/H/H2, K117N/N2, A146T/V
PCR method: PlexPrime/PlexZyme Cq curve
Detection limit: 1–5%
Turnaround time: 2 hours (approx.)
No relevant test availableAmpliSeq Cancer Hotspot Panel v224
Coverage* A11T, G12N/R/C/S/D/A/V/G, G13R/C/S/V/D/A/G, A18T, G60E, Q61L/K/R/E/P/L/R/H/Q, S65C, A146T
Detection limit: see above
Turnaround time: single day
  • Turnaround times and detection limits (analytical sensitivity) are given as quoted by the manufacturers. The Ion PGM (NGS) panel is that described on the Thermo Fisher website.24

  • *Coverage given is for the codon changes that are likely to be relevant in CRC.

  • †Terminating codon notation.

  • CRC, colorectal carcinoma; NGS, next-generation sequencing; PCR, polymerase chain reaction; PGM, Personal Genome Machine; v2, version 2.