Table 2.

Evaluation of IDH1 status received from non-reference center (MLPA), with 3 molecular methods (NGS, qPCR, Sanger) received from reference oncology center

Patient ID	Non-reference center		Reference oncology center
	Histopathological diagnosis	IDH1 status MLPA	IDH1 status NGS	First verification qPCRa	Second verification Sanger	Protein change	Concordance in IDH1 status
12	A3	wt	wt	wt	wt	p=	Yes
13	A2	wt	Pathogenic	Mutant	Mutant	R132Lb	No*
19	A2	Mutant	Pathogenic	Mutant	Mutant	R132Hc	Yes
34	A2	Mutant	Pathogenic	Mutant	Mutant	R132Hc	Yes

A2/3 astrocytoma grade 2/3, IDH1 isocitrate dehydrogenase 1, MLPA multiplex ligation-dependent probe amplification to detect 4 mutant variants (R132C, R132H, R172K, R172M), NGS next-generation sequencing, p= no protein change has been detected in analyzed sample, qPCR quantitative polymerase chain reaction, wt wildtype

^aIDH1/2 Mutation Analysis Kit (EntroGen) is a screening qPCR test for the detection of the most common mutations in codon 132 and 100 of the IDH1 gene and in codon 172 and 140 of the IDH2 gene. IDH1: Codon 132 (the test cannot distinguish between the following mutations: c.395G>A; c.394C>T; c.394C>A; c.394C>G; c.395G>T; c.395G>C; c.394_395delinsGT); codon 100 (c.299G>A)

^bNM_005896.2:c.395G>T

^cNM_005896.2:c.395G>A

^*MLPA test does not detect R132L variant