Table 1.

Comparison of different molecular genetic tests for CALR mutations detection in patients with suspected myeloproliferative neoplasm.

Method	Advantage	Critical Remarks	Sensitivity	Reference
Sanger sequencing	Known and unknown genetic variant detection.	Low sensitivity; Not quantitative; Moderate cost.	10 to 20%	[9,10,60]
PCR and fragment analysis	Known and unknown genetic variant detection; Qualitative and quantitative; Simple to perform; Low cost and rapid.	Moderate to low sensitivity.; Preferential amplification of shorter amplicons may lead to over- or underestimation of the mutant allele burden; Sanger sequencing needed for correctly genotype the CALR variants.	1 to 10%	[9,60,61,72,80,85]
High-resolution Melt	Known and unknown genetic variant detection; Simple to perform; Low cost and rapid.	Moderate to low sensitivity; Not quantitative; Sanger sequencing needed for correctly genotype the CALR variants.	1 to 5%	[60,66,67,68]
Quantitative PCR (real-time PCR) (qPCR)	High sensitivity; Quantitative; Rapid.	Detects only target genetic variants; Moderate cost.	0.01 to 1%	[60,62,71,86]
Digital PCR	High sensitivity; Quantitative; Rapid.	Detects only target genetic variants; Moderate cost.	0.01 to 1%	[28,72,86,87,88]
NGS	Known and unknown genetic variant detection; Simultaneous screening of multiple genes in multiple samples.	Complex genetic variants and large indels need in some instances confirmation by alternate molecular genetic methods; Complex workflow and result interpretation; Moderate to high cost.	1 to 5%	[60,75,80]

Abbreviations: PCR, polymerase chain reaction; NGS, next generation sequencing.