Assay- Qualitative detection of mutations in EGFR gene
Kit used – “EasyPGX® ready EGFR” kit detects 85 mutations of the oncogene EGFR
Method used – Real-Time PCR
Sample type: FFPE (tumor cell content >20 %)
List of detectable mutations:
- EGFR G719x: the assay detects 4 mutations of codon 719 but does not distinguish between them
- EGFR T790M: the assay detects the T790M mutation
- EGFR S768I: the assay detects the S768I mutation
- EGFR ex20ins: the assay detects 11 mutations/insertions of the exon 20 of EGFR but does not distinguish between them
- EGFR L858R: the assay detects four L858R mutations
- EGFR L861Q: the assay detects the L861Q mutation
- EGFR ex19del: the assay detects the most frequent 64 deletions of the nucleotides of the exon 19 of EGFR but does not distinguish between them
*(not distinguishable between them)
| EGFR G719x * | EGFR T790M | EGFR S768I | EGFR ex20ins * | EGFR L858R * | EGFR L861Q | EGFR ex19del * |
| ▪ G719S (2155G>A) | ▪ T790M (2369C>T) | ▪ S768I (2303G>T) | ▪ V769_D770insASV (2307_2308insGCCAGCGTG)* | ▪ L858R (2573T>G)* | ▪ L861Q (2582T>A) | ▪ E746_A750del (2235_2249del15) |
| ▪ G719C (2155G>T) | ▪ D770_N771insG (2310_2311insGGT)* | ▪ L858R (2573_2574TG>GA) | ▪ E746_A750del (2236_2250del15) | |||
| ▪ G719A (2156G>C) | ▪ H773_V774insH (2319_2320insCAC)* | ▪ L858R (2573_2574TG>GT) | ▪ L747_P753>S (2240_2257del18) | |||
| ▪ G719fs*29 (2156G>-) | ▪ V769_D770insASV# (2307_2308insGCCAGCGTG) | ▪ L747_A750>P (2239_2248TTAAGAGAAG>C) | ||||
| ▪ V769_D770insASV (2296_2297insTGGCCAGCG)* ** | ▪ E746_S752>V (2237_2255>T) | |||||
| ▪ V769_D770insASV (2308_2309insCCAGCGTGG) | ▪ L747_T751del (2240_2254del15) | |||||
| ▪ V769_D770insASV (2309_2310AC>CCAGCGTGGAT)* ** | ▪ L747_S752del (2239_2256del18) | |||||
| ▪ D770_N771>AGG (2309_2312ACAA>CTGGTGG)* ** | ▪ E746_T751>A (2237_2251del15) | |||||
| ▪ N771>GY (2311A>GGTT)* ** | ▪ L747_T751del (2239_2253del15) | |||||
| ▪ N771>SH (2311_2312insGTC)* ** | ▪ L747_T751>P (2239_2251>C) | |||||
| ▪ N771>SGH (2311_2312insGTGGCC) | ▪ L747_E749del (2239_2247del9) | |||||
| ▪ E746_E749del (2235_2246del12) | ||||||
| ▪ L747_P753>Q (2239_2258>CA) | ||||||
| ▪ L747_T751>S (2240_2251del12) | ||||||
| ▪ E746_S752>A (2237_2254del18) | ||||||
| ▪ L747_A750>P (2238_2248>GC) | ||||||
| ▪ E746_S752>D (2238_2255del18) | ||||||
| ▪ E746_T751>I (2235_2252>AAT) | ||||||
| ▪ L747_T751>Q (2238_2252>GCA) | ||||||
| ▪ E746_T751del (2236_2253del18) | ||||||
| ▪ K745_E746insIPVAIK (2232_2233ins18) ** | ||||||
| ▪ K745_E746insIPVAIK (2234_2235ins18) ** | ||||||
| ▪ K745_E746insTPVAIK (2234_2235ins18) ** | ||||||
| ▪ E746_E749delELRE (2235_2246del12) ** | ||||||
| ▪ E746_A750>IP (2235_2248>AATTC) ** | ||||||
| ▪ E746_T751>IP (2235_2251>AG) ** | ||||||
| ▪ E746_T751delELREAT (2235_2252del18) ** | ||||||
| ▪ E746_S752>I (2235_2255>AAT) ** | ||||||
| ▪ E746delE (2234_2236delAGG) ** | ||||||
| ▪ E746_T751>S (2236_2251>T) ** | ||||||
| ▪ E746_A750>AP (2237_2248>CAC) ** | ||||||
| ▪ K745_E746insVPVAIK (2236_2237ins18) ** | ||||||
| ▪ E746_S752>V (2235_2255>GGT) ** | ||||||
| ▪ E746_T751>VP (2237_2251>TTC) ** | ||||||
| ▪ E746_T751>VA (2237_2253>TTGCT) ** | ||||||
| ▪ E746_S752>V (2237_2256>TT) ** | ||||||
| ▪ E746V (2237_2238AA>TT) ** | ||||||
| ▪ E746_T751>VP (2237_2253>TTCCT) ** | ||||||
| ▪ E746_P753>VQ (2237_2258>TTCA) ** | ||||||
| ▪ E746_A750>DP (2238_2249>TCC) ** | ||||||
| ▪ L747_T751>P (2238_2251>GC) ** | ||||||
| ▪ L747_S752>QH (2238_2255>GCAACA) ** | ||||||
| ▪ L747_S752>Q (2238_2256>GCAA) ** | ||||||
| ▪ L747_T751>A (2239_2253>GCT) ** | ||||||
| ▪ L747_K754delLREATSPK (2239_2262del24) ** | ||||||
| ▪ L747_A755>AN (2239_2264>GCCAA) ** | ||||||
| ▪ L747_K754>ANKG (2239_2262>GCCAACAAGGGA) ** | ||||||
| ▪ L747P (2239_2240TT>CC) ** | ||||||
| ▪ L747_A750>P (2239_2250>CCA) ** | ||||||
| ▪ L747_T751>PT (2239_2253>CCAACG) ** | ||||||
| ▪ L747_T751delLREAT (2238_2252del15) ** | ||||||
| ▪ L747_P753>S (2239_2257>T) ** | ||||||
| ▪ L747S (2240T>C) ** | ||||||
| ▪ L747_K754>ST (2240_2261>CGAC) ** | ||||||
| ▪ L747_A755>SKG (2240_2264>CGAAAGG) ** | ||||||
| ▪ L747_T751>Q (2239_2253>CAA) ** | ||||||
| ▪ L747_T751>Q (2239_2252>CA) ** | ||||||
| ▪ L747_S752>Q (2239_2256>CAA) ** | ||||||
| ▪ L747_S752>QH (2239_2256>CAACAT) ** | ||||||
| ▪ K745_E746insIPVAIK (2235_2236ins18) ** | ||||||
| ▪ L747_E749delLRE (2236_2244delGAATTAAGA) ** | ||||||
| ▪ L747_A750>P (2239_2250>CCG) ** | ||||||
| ▪ L747_A750>S (2240_2248del9) ** | ||||||
| ▪ L747_P753>Q (2239_2259>CAA) ** |
Limitations
| Assay | LOD C95 (at medium DNA input concentration) |
| G719x | 5.0% |
| T790M | 1.0% |
| S768I | 0.5% |
| ex20ins | 5.0% |
| L858R | 1.0% |
| L861Q | 1.0% |
| ex19del | 0.5%
|
Disclaimer
This assay can detect only EGFR mutations mentioned above.
This assay’s accuracy depends on the sample’s quality, tumor content, and allele frequency.
