Table 4.

Comparison of characteristics of methods for determining TERTp mutations.

The Method	The Amount of Material Required	Number of Mutations Determined Simultaneously	Sensitivity of the Method	Method Specificity	Cost of Analysis	Technical Complexity	Time of Analysis	Pros, Cons and Limitations
ddPCR	Low (1–5 ng of DNA)	One	1%	High	High	High	During the working day	+: Low amount of the DNA template; absolute quantification; resistance to PCR inhibitors −: High cost of assays; need for well-trained staff; higher contamination risk; limited and defined targets
Multiplexed dPCR	Low (1–5 ng of DNA)	Several	1–2%	High	Medium	Medium	During the working day	+: Low amount of the DNA template; feasibility; absolute quantification; resistance to PCR inhibitors; user-friendly system −: Considerable cost of analyses; high contamination risk; limited and defined targets
dPCR	Low (1–5 ng of DNA)	One	1%	High	Medium	Medium	During the working day	+: Low amount of the DNA template; feasibility; absolute quantification; resistance to PCR inhibitors; user-friendly system −: Considerable cost of analyses; single-target method; high contamination risk; limited and defined targets
NGS mutation panel	Moderate (5–10 ng of DNA)	A lot of	1 to 5%	High	High	Very high	In a few days	+: Satisfactory estimate of MAF; availability of diverse commercial tests; possibility of detecting large diversity of targets including unpredictable mutations and allelic forms −: High cost of assays; time-consuming method
Nanopore sequencing	Very low (1 ng of DNA)	A lot of	1%	High	High	High	Within 6 h	+: Possibility of detecting large diversity of targets including unpredictable mutations and allelic forms −: High error rates; high cost
Sanger sequencing	High (10 ng of DNA)	Several	10 to 20%	Medium	Low	Low	Within 4–6 h	+: Low cost −: High error level; high Limit of Detection; quantitative tests are problematic