Association between calcium sensing receptor gene polymorphisms and chronic pancreatitis in a US population: Role of serine protease inhibitor Kazal 1type and alcohol

Table 1 Polymerase chain reaction primer pairs, magnesium concentration and annealing temperatures used for genetic analysis of the CASR gene

Scanning region	Forward and reverse primer sequences	MgCl2 (mmol/L)	Annealing temperature (°C)
Exon 2	5’-ACCACCCACATTACAAGTC-3’	2.5	55
	5’-GCTTTTCTCCAACCACTCAG-3’
Exon 3	5’-ATGAAGCCAGAGAGTAGTAAC-3’	2.5	58
	5’-TAAACCGTATGGCTATTGGG-3’
Exon 4a	5’-GCTTTTCCTTACCCTTTCTTTCATC-3’	2	58
	5’-ATCACCTCTACCACATGCTG-3’
Exon 4b	5’-CAGATCTTGAGCCCCTCATC-3’	2	59
	5’-GCAGCCCAACTCTGCTTTAT-3’
Exon 5	5’-TGGGGCTTGTACTCATTCTT-3’	1.5	59
	5’-CTGGTTTTCTGATGGACAGC-3’
Exon 7a	5’-CACACAATAACTCACTCTTCAC-3’	2.0	61
	5’-CAGAGGAAAACCAGCAGGAAC-3’
Exon 7b	5’-AAAACCAACCGTGTCCTCCTG-3’	1.0	53
	5’-ATGGCAATCACCTCTACGGC-3’
Exon 7c	5’-GCTCATCTTCTTCATCGTCTGG-3’	1.0	58
	5’-CGTATCGCTGCTTTTCTGGG-3’
Exon 7d	5’-CCCAGCAAGAGCAGCAG-3’	1.0	58
	5’-ACAACTCTTCAGGGTCCTCC-3’

Table 2 Participant characteristics

Demographic	CP (n = 219)	RAP (n = 115)	Controls (n = 305)
Age, mean (SD)	45.3 (18.1)	46.1 (16.2)	54.7 (14.5)
Race, % White	91	91	94
Sex (M/F)	125/94	49/66	121/184
Alcohol drinking pattern (%)
Abstainers	45 (22.5)	30 (27)	72 (25)
Light	38 (19)	26 (23)	83 (28)
Moderate	34 (17)	22 (20)	56 (19)
Heavy	39 (19.5)	20 (18)	58 (20)
Very heavy	44 (22)	13 (12)	23 (8)

CP: Chronic pancreatitis; RAP: Recurrent acute pancreatitis; SD: Standard deviation. Abstainers: No alcohol use or < 20 drinks in lifetime; Light: < 3 drinks/week; Moderate: 4-7 drinks/week for females, 4-14 drinks/week for males; Heavy; 8-34 drinks/week for females, 15-34 drinks/week for males; Very heavy: > 35 drinks/week.

Table 3 Genotype analysis of CASR R990G polymorphism in patients and controls

	Patients	Controls	P1	OR (95% CI)
CP patients vs controls without SPINK1 N34S (%)
AA	140 (82)	255 (90)
AG	31 (18)	28 (10)
GG	1 (1)	1 (0.5)
AA vs AG/GG			0.015	2.01 (1.12-3.59)
RAP patients vs controls without SPINK1 N34S (%)
AA	93 (89)	255 (90)
AG	10 (9)	28 (10)
GG	2 (2)	1 (0.5)
AA vs AG/GG			0.712	1.28 (0.67-2.47)
SPINK1 N34S positive CP patients vs controls (%)
AA	39 (83)	20 (95)
AG	8 (17)	1 (5)
GG	0	0
AA vs AG/GG			0.255	4.1 (0.48-35.14)

CP: Chronic pancreatitis; RAP: Recurrent acute pancreatitis. SPINK1: Serine protease Kazal type 1 gene.

¹Fisher exact test.

Table 4 CASR genotype comparison for R990G polymorphism in CP patients and controls with similar alcohol drinking patterns

	Patients	Controls	P1	OR (95% CI)
A/L Alcohol CP patients vs controls (%)
AA	69 (83)	136 (88)
AG	13 (16)	18 (11)
GG	1 (1)	1 (1)
AA vs AG/GG			0.332	1.45 (0.69-3.07)
M/H Alcohol CP patients vs controls (%)
AA	59 (81)	106 (93)
AG	14 (19)	8 (7)
GG	0	0
AA vs AG/GG			0.018	3.12 (1.14-9.13)
VH Alcohol CP patients vs controls (%)
AA	37 (84)	21 (91)
AG	7 (16)	2 (9)
GG	0	0
AA vs AG/GG			0.708	1.99 (0.38-0.45)

A: Abstainer; L: Light; M: Moderate; H: Heavy; VH: Very heavy. Alcohol categories: Abstainers: No alcohol use or < 20 drinks in lifetime; Light drinkers: < 3 drinks/week; Moderate drinkers: 4-7 drinks/week for females, 4-14 drinks/week for males; Heavy drinkers: 8-34 drinks/week for females, 15-34 drinks/week for males; Very heavy drinkers: > 35 drinks/week.

¹Fisher’s exact test.