A comparison of Direct sequencing, Pyrosequencing, High resolution melting analysis, TheraScreen DxS, and the K-ras StripAssay for detecting KRAS mutations in non small cell lung carcinomas

Table 5 Summary of the properties of the different methods

	Sanger sequencing	Pyrosequencing	TheraScreen DxS	StripAssay	HRM
CE mark	no	no	yes	yes	no	CE mark
Limit of detection*	25-30 %*	5-10 %*	1 %	below 1 %	5-10 %*	Limit of detection*
Turnaround time	2-3 days	2 days	1/2 day	1 day	1/2 day	Turnaround time
Ease of interpretation	easy	easy	easy	medium	difficult	Ease of interpretation
Technician time	6 hrs	4 hrs	2 hrs	5 hrs	2 hrs	Technician time
Amount of input DNA	1 reaction	1 reaction	8 reactions	1 reaction	1 reaction	Amount of input DNA
Detection of rare mutations	Yes – can detect any mutation located between the primers.	Yes – can detect any mutation within the short sequencing fragments.	No – can only detect 7 specific mutations.	No – can only detect 10 specific mutations.	Yes – can detect some mutations located between the primers.	Detection of rare mutations
Reagent cost	20 €	40 €	120 €	60 €	4 €	Reagent cost
Special equipment required	Sequencer 70 000 €	Pyrosequencer 150 000 €	Real time PCR cycler 30 000 €	PCR cycler 7 500 €	HRM Real time PCR cycler 75 000 €	Special equipment required

ISSN: 1756-9966