Advantages and disadvantages of the main methods currently available for BCR::ABL1 KD mutation testing
| Method . | Lower limit of detection . | Mutation-specific? . | Advantages . | Disadvantages . |
|---|---|---|---|---|
| Sanger sequencing | 10%-20% | N | Enables TKD-wide screening; easy workflow and data analysis; relatively short turnaround time | Poorly sensitive |
| Mass spectrometry | 0.05%-0.5% | Y | Accurate; highly sensitive | Not widely available; high throughput; longer turnaround time; no commercial kits available; can be implemented for a limited number of mutations; difficult identification of the T315L and T315M mutations (that are caused by a double nucleotide substitution) |
| NGS | 1%-5% | N | Enables TKD-wide screening; sensitive | High throughput; longer turnaround time; requires specialized personnel and bioinformatic competences; relatively high error rate; no commercial kits available |
| ddPCR | 0.1%-0.5% | Y | Accurate; highly sensitive; relatively easy workflow and data analysis; short turnaround time | Can be implemented for a limited number of mutations |
| Method . | Lower limit of detection . | Mutation-specific? . | Advantages . | Disadvantages . |
|---|---|---|---|---|
| Sanger sequencing | 10%-20% | N | Enables TKD-wide screening; easy workflow and data analysis; relatively short turnaround time | Poorly sensitive |
| Mass spectrometry | 0.05%-0.5% | Y | Accurate; highly sensitive | Not widely available; high throughput; longer turnaround time; no commercial kits available; can be implemented for a limited number of mutations; difficult identification of the T315L and T315M mutations (that are caused by a double nucleotide substitution) |
| NGS | 1%-5% | N | Enables TKD-wide screening; sensitive | High throughput; longer turnaround time; requires specialized personnel and bioinformatic competences; relatively high error rate; no commercial kits available |
| ddPCR | 0.1%-0.5% | Y | Accurate; highly sensitive; relatively easy workflow and data analysis; short turnaround time | Can be implemented for a limited number of mutations |
ddPCR, droplet digital polymerase chain reaction; N, no; NGS, next generation sequencing; TKD, tyrosine kinase domain; Y, yes.