A Pilot Study of ANXA2, MED12, CALM1 and MAPK1 Gene Variants in Primary Hyperparathyroidism

Statistical analysis of rs7170178 (ANXA2 gene) and rs12885713 (CALM1 gene) variants between PHPT patients and controls_

Gene: Genotypes of the studied variant	Patients	Controls	Statistical model	OR (95%CI)	p-Value	HWE in control group (p-value)
ANXA2: rs7170178
AA	18	22	Additive (AA vs. AG vs. GG)		0.45	0.78
AG	29	23
GG	3	5
AA	18	22	Homozygous (GG vs. AA)	0.73 (0.15-3.49)	1
GG	3	5
AA	18	22	Heterozygous (AG vs. AA)	1.54 (0.67-3.53)	0.31
AG	29	23
AA	18	22	Dominant (AG+GG vs. AA)	1.4 (0.62-3.11)	0.41
AG+GG	32	28
AA+AG	47	45	Recessive (GG vs. AA+AG)	0.57 (0.13-2.55)	0.71
GG	3	5
A	65	67	Allelic (G vs. A)	1.09 (0.61-1.96)	0.76
G	35	33
CALM1: rs12885713
CC	20	14	Additive (CC vs. CT vs. TT)		0.39	0.98
CT	19	25
TT	11	11
CC	20	14	Homozygous (TT vs. CC)	0.7 (0.24-2.06)	0.52
TT	11	11
CC	20	14	Heterozygous (CT vs. CC)	0.53 (0.21-1.32)	0.17
CT	19	25
CC	20	14	Dominant (CT+TT vs. CC)	0.58 (0.25-1.35)	0.21
CT+TT	30	36
CC+CT	39	39	Recessive (TT vs. CC+CT)	1 (0.38-2.57)	1
TT	11	11
C	59	53	Allelic (T vs. C)	0.78 (0.45-1.37)	0.39
T	41	47

Primer sequences, restriction enzymes, and restriction digestion patterns were used for genotyping of the studied genetic variants_

Gene	Variant	Primer sequence (5′-> 3′)	Amplicon size	Restriction Enzyme	Restriction digestion pattern
ANXA2	rs7170178	F: 5’-TTCACAGCAGTTCAAAATAC-3’R: 5’-CTGGGTTTCCAGAGATGGAA-3’	550bp	HpyCH4V	G: 338bp/106bp/72bp/34bpA: 195bp/143bp/106bp/72bp/34bp
	rs17191344	F: 5’-TGCAAACAGGCGCCACTAAA-3’R: 5’-CAGACATGAGGCCAAGGAACT-3’	280bp	Hpy99I	A: 94bp/186bpG: 139bp/94bp/47bp
	rs11633032	F: 5’-CAACAAGCATGGGGTTGC-3’R: 5’-GTTGACATTTGCCCTTCGCTT-3’	131bp	BceAI	G: 123bp/8bpA: 131bp
MED12	rs1057519912	F: 5’-ACAAGCCTACAGTAGGAATCC-3’R: 5’-TGGCACCACTCCCTTCCTAC-3’	143bp	Smll	C: 94bp/49bpG/T: 143bp
CALM1	rs12885713	F: 5’-GGGATACGGCGCACCATATAT-3’R: 5’-GGTACCTCCGATGCCGCTG-3’	179bp	HpyCH4V	T: 144bp/35bpC: 179bp
MAPK1	rs1057519911	F: 5’-TGGCTGATCTATGTCCCTGA-3’R: 5’-CACACAAGAGGATTGAAGTAG-3’	122bp	MnlI	C: 63bp/59bpT: 122bp