Analysis of the human Atox 1 homologue in Wilson patients

Table 1 Oligonucleotide primers used to amplify Atox1

Exon	Sequence	Fragment size
Exon 1-Forward primer	5’-GGAGTGGGAGGGGCCTCCGGGACC-3’	273 bp
Exon 1-Reverse primer	5’-GTAAGCTAGGGGACAACAGCGGCTC-3’	273 bp
Exon 2 - Forward primer	5’-GCACTGTGTGGGGGTCACTCTACAG-3’	320 bp
Exon 2-Reverse primer	5’-GTGAGGATTAAATGATGTGATTCAC-3’	320 bp
Exon 3-Forward primer	5’-GAACTCTTTCTTGCTGTAACTGGGAG-3’	344 bp
Exon 3-Reverse primer	5’-GAGGGCTCTCCCGCTCCACTCAAG-3’	344 bp
Exon 4-Forward primer	5’-TGCAATGTCGCTATGTCCACACCA-3’	326 bp
Exon 4-Reverse primer	5’-GATCACACAGCAAAGAATCAGAATC-3’	326 bp

Table 2 Atox1 gene analysis in Wilson disease patients

		Distribution of the variations detected in the Atox1-gene among the ATP7B-genotypes
Mutations in the ATP7B-gene	No. of patients	5’UTR -99 T>C		5’UTR -68 C>T		No variation within the sequence
		Heterozygous	Homozygous	Heterozygous	Homozygous
H1096Q/H1096Q	22	6	4	-	-	12
R1041W/R1041W	2				-	2
G12810N/G12810N	1				-	1
G1266R/G1266R	1	1			-
G710A/G710A	1					1
K844k-fs/K844k-fs	1					1
W779X/W779X	1		1
D765N/Y741X	2		1			1
3400 Del C/2299InsC	1					1
3400 Del C/H1096Q	1	1
3400 Del C/G982V	1	1
3400 Del C/W779X	1					1
H1096Q/M769V	1	1
m.n.d./m.n.d.	11	7		2		4
H1096Q/m.n.d.	8	3	1			4
W 778 X/m.n.d.	1	1
W779X/m.n.d.	2		1			1
D765N/m.n.d.	2					2
G710S/m.n.d.	1		1
M769V/m.n.d.	1					1
2299 InsC/m.n.d.	1	1
Total	63	22	9	2	0	32

m.n.d.: Mutation not detected.

Table 3 Analysis of average ages of initial manifestation with regard to the variations detected in the Atox1 gene

Atox1-variation	Average age of initial manifestation	n of patiens	Standard deviation
No variation within the sequence	23.68	31	12.993
Variation within the sequence detected	18.88	25	9.909
Total	21.54	56	11.863

Table 4 Statistical data for Analysis of age of initial manifestation

	Not standardized coefficients		Standardized coefficients
Model	B	Standard error	Beta	T	Significance
ATP7B	4.042	1.970	0.272	2.052	0.045
Atox1	-5.312	3.079	-0.225	-1.725	0.091