Development and validation of a multiplex-PCR assay for X-linked intellectual disability

Table 1 Validation cohort results

n	FMR1CGG	AFF2CCG	ARX(bp)	n	FMR1CGG	AFF2CCG	ARX(bp)	n	FMR1(CGG)	AFF2(CCG)	ARX(bp)	Obtained/confirmed by Southern bloting
1	19	13	380	3	30	11	380	1	33	22	380
1^a	19	47	380	4	30	13	380	1	35	34	380
2	20	14	380	1^b	30	14	404	2	36	14	380
1	20	15	380	14	30	14	380	1	37	23	380
1	21	14	380	1	30	15	380	1	41	23	380
1	21	15	380	1	30	16	380	1	42	23	380
1	21	23	380	4	30	17	380	1	65	14	380	~70
3	23	14	380	2	30	18	380	1	71	14	380	~70
1	23	16	380	1	30	19	380	1	75	23	380	~80
2	23	17	380	2	30	20	380	1	80	22	380	~80
2	23	19	380	1	30	22	380	1	85	14	380	~80
1	23	20	380	1	30	24	380	1	100	15	380	~100
1	24	14	380	2	30	27	380	1	110	14	380	~100
1	27	14	380	1	31	14	380	1	120	15	380	~120
1	28	20	380	2	31	17	380	1	- (150)	21	380	~150
1	29	8	380	1	31	20	380	1	-	17	380	~165
3	29	14	380	1	31	21	380	1	-	16	380	~250
1	29	15	380	1	31	30	380	1^c	51	22	380	~350
1	29	16	380	2	32	14	380	1	-	17	380	~350
1	29	9	380	1	32	15	380	1	-	14	380	~400
4	29	17	380	1	32	19	380	1	-	13	380	~650

Number of triplets obtained for FMR1 and AFF2 genes, and ARX ex2p fragment size, in 100 male samples. The order is according to FMR1 allele sizes. Number of triplets obtained for FMR1 and AFF2 genes, and ARX ex2p fragment size, in 100 male samples, ordered according to FMR1 allele sizes. [n] Number of samples per allele profile; ^a AFF2 intermediate size allele; ^bcontrol for the ARX c.429_452dup; ^cFXS sample with mosaicism for an intermediate-size allele and a full mutation; Premutations clearly amplified by the multiplex-PCR and additionally confirmed by sequencing are bold highlighted; [−] not (reproducibly) amplified.