Table 1.

Comparison among various detection methods for gene mutation analysis

	Sanger sequencing	Pyrosequencing	Real-time PCR	PNA-clamp assay	Next generation sequencing
Advantage	Gold standard	More rapid and sensitive than Sanger sequencing	Simple and fast	Simple and fast	High-throughput
Instrument	Ubiquitous	Not ubiquitous	Depending on kit	Ubiquitous	Costly equipment
Sensitivity (%)	10–20	5	1	0.1	1
Method	Labor-intensive, time-consuming	Convenient, closed system	Closed system, one-step process	Closed system, one-step process	Time-consuming
Mutant allele quantity (%)	Unmeasurable	Measurable	Unmeasurable	Unmeasurable	Measurable
Detect all or new mutation	Yes	No	No	No	Yes
Detect specific mutation	Yes	Yes	No	No	Yes
Data interpretation	Subjective	Less subjective, but complex	Simple and easy	Simple and easy	Complicated (need statistics)

PCR, polymerase chain reaction; PNA, peptide nucleic acid.