TABLE 11.
Comparison of Sanger Sequencing to Other Methods for the Detection of EGFR Mutationsa
| Incidence of EGFR Mutations, % |
||||||||
|---|---|---|---|---|---|---|---|---|
| Sanger Sequencing Versus Other Methods | Concordance of Methods, % | n (N) | Seq−/Other+ | Seq+/Other− | Sanger Sequencing | Other Method | RR (95% CI) | P Value |
| PCR-based mutation detection | ||||||||
| Allele-specific PCR/ARMS130 | 73 | 1 (83) | 18 | 4 | 16 | 33 | 0.48 (0.27–0.87) | .01 |
| Real-time PCR119,304 | 97 | 2 (102) | 2 | 1 | 26 | 27 | 0.94 (0.60–1.46) | .78 |
| Cycleave PCR131,b | 95 | 1 (195) | 1 | 8 | 40 | 36 | 1.10 (0.85–1.41) | .47 |
| Post-PCR mutation detection | ||||||||
| Capillary electrophoresis296,304 | 98 | 2 (61) | 1 | 0 | 16 | 18 | 0.91 (0.42–2.01) | .82 |
| Restriction fragment length polymorphism133 | 99 | 1 (109) | 1 | 0 | 33 | 34 | 0.97 (0.67–1.41) | .89 |
| INVADER305,c | 86 | 1 (42) | 5 | 1 | 43 | 52 | 0.82 (0.52–1.29) | .39 |
| Pyrosequencing159,306 | 96 | 3 (140) | 6 | 0 | 16 | 20 | 0.78 (0.49–1.25) | .30 |
| Mutation scanning | ||||||||
| Denaturing HPLC139,307 | 94 | 2 (196) | 12 | 0 | 20 | 27 | 0.66 (0.27–1.63) | .37 |
| Single-stranded conformational polymorphism45 | 98 | 1 (375) | 8 | 0 | 8 | 10 | 0.79 (0.51–1.25) | .32 |
| High-resolution melting analysis211,135,308 | 83 | 3 (321) | 54 | 0 | 36 | 53 | 0.70 (0.46–1.06) | .09 |
| Loop-hybrid mobility shift assay309 | 100 | 1 (43) | 0 | 0 | 26 | 26 | 1.00 (0.49–2.06) | >.99 |
| Mutant enrichment | ||||||||
| Peptide nucleic acid/locked nucleic acid amplification301,310 | 96 | 2 (150) | 4 | 2 | 11 | 12 | 0.91 (0.49–1.67) | .75 |
| COLD PCR138 | 100 | 1 (126) | 0 | 0 | 10 | 10 | 1.00 (0.48–2.07) | >.99 |
| Smart Amplification Process120,132,301 | 86 | 4 (220) | 30 | 0 | 20 | 34 | 0.58 (0.44–0.77) | <.001 |
Abbreviations: ARMS, amplification refractory mutation system; CI, confidence interval; COLD, coamplification at low denaturation temperature; HPLC, high-performance liquid chromatography; n, number of studies; N, number of patients; PCR, polymerase chain reaction; RR, relative risk, Mantel-Haenszel random effects model, (95% CI); Seq−, negative by Sanger sequencing; Seq+, positive by Sanger sequencing.
The variation in EGFR mutation rate between rows may reflect studies performed in different patient populations (Asian versus non-Asian). No statistical comparisons were performed between rows.
Cycleave; Takura Bio, Otsu, Shiga, Japan.
Invader; Hologic, Madison, WI.