Association between NOD2/CARD15 gene polymorphisms and Crohn's disease in Chinese Zhuang patients

Table 1 Polymerase chain reaction primers, annealing temperatures and polymerase chain reaction fragment sizes

Mutation	Primer	Annealing temperature (°C)	PCR fragment size (bp)
P268S	F-TGCCTCTTCTTCTGCCTTCC	60	422
	R-AGTAGAGTCCGCACAGAGAG
JW1	F-TGCAGTTTTCTTGGGGAGAT	59	220
	R-TGTACCTGATCCAGCCCAAT
N852S	F-CTGTTTGCATGATGGGGGG	55	151
	R-CAGCCGTCAGTCAATTTGTAG

PCR: Polymerase chain reaction.

Table 2 Enzymes and gene polymorphism analysis

Mutation	Base change	Enzyme	Restriction fragment size (bp)
			Wild-type	Mutant
P268S	C→T	BamHI	422	Heterozygote 422 + 247 + 175
				Homozygote 247 + 175
JW1	C→T	XhoI	125 + 95	Heterozygote 220 + 125 + 95
				Homozygote 220
N852S	A→G	AluI	151	Heterozygote 151 + 129 + 22
				Homozygote 129 + 22

Table 3 Distribution of genotype and allele frequencies of mutations in Crohn’s disease and ulcerative colitis patients compared with healthy controls in the Guangxi Zhuang population n (%)

Mutant	Genotype	Allele	Control	CD		UC
					P value		P value
P268S					aP1		NS1
	CC		71 (98.6)	42 (87.5)		54 (100.0)
	CT		1 (1.4)	6 (12.5)		0 (0.0)
	TT		0 (0.0)	0 (0.0)		0 (0.0)
		T	1 (0.7)	6 (6.2)	cP2	0 (0.0)	NS2
JW1					NS1		NS1
	CC		72 (100.0)	48 (100.0)		54 (100.0)
	CT		0 (0.0)	0 (0.0)		0 (0.0)
	TT		0 (0.0)	0 (0.0)		0 (0.0)
		T	0 (0.0)	0 (0.0)	NS2	0 (0.0)	NS2
N852S					NS1		NS1
	AA		72 (100.0)	48 (100.0)		54 (100.0)
	AG		0 (0.0)	0 (0.0)		0 (0.0)
	GG		0 (0.0)	0 (0.0)		0 (0.0)
		G	0 (0.0)	0 (0.0)	NS2	0 (0.0)	NS2

¹Comparisons of genotype frequencies;

²Comparisons of allele frequencies.

^aP < 0.05 vs control using Fisher's exact test;

^cP < 0.05 vs control using Fisher's exact test. CD: Crohn’s disease; UC: Ulcerative colitis; NS: No significance.

Table 4 Distribution of genotype and allele frequencies of mutations in Crohn’s disease and ulcerative colitis patients compared with healthy controls in the Guangxi Han population n (%)

Mutant	Genotype	Allele	Control	CD		UC
					P value		P value
P268S					bP1		NS1
	CC		78 (100.0)	46 (92.0)		48 (96.0)
	CT		0 (0.0)	4 (8.0)		2 (4.0)
	TT		0 (0.0)	0 (0.0)		0 (0.0)
		T	0 (0.0)	4 (4.0)	dP2	2 (2.0)	NS2
JW1					NS1		NS1
	CC		78 (100.0)	50 (100.0)		50 (100.0)
	CT		0 (0.0)	0 (0.0)		0 (0.0)
	TT		0 (0.0)	0 (0.0)		0 (0.0)
		T	0 (0.0)	0 (0.0)	NS2	0 (0.0)	NS2
N852S					NS1		NS1
	AA		78 (100.0)	50 (100.0)		50 (100.0)
	AG		0 (0.0)	0 (0.0)		0 (0.0)
	GG		0 (0.0)	0 (0.0)		0 (0.0)
		G	0 (0.0)	0 (0.0)	NS2	0 (0.0)	NS2

¹Comparisons of genotype frequencies;

²Comparisons of allele frequencies.

^bP < 0.05 vs control using Fisher's exact test;

^dP < 0.05 vs control using Fisher's exact test. CD: Crohn’s disease; UC: Ulcerative colitis; NS: No significance.

Table 5 Clinical characteristics of Crohn’s disease patients in Guangxi with and without P268S mutations

Phenotype	n	P268S+	P268S-	P value
Age of onset				aP
≤ 40 years	45	8 (17.8)	37 (82.2)
> 40 years	53	2 (3.8)	51 (96.2)
Location				NS
Ileum	58	6 (10.3)	52 (89.7)
Colon/ileocolon	40	4 (10.0)	36 (90.0)
Gender				NS
Male	51	7 (13.7)	44 (86.3)
Female	47	3 (6.4)	44 (93.6)
Ethnic groups				NS
Han	50	4 (8.0)	46 (92.0)
Zhuang	48	6 (12.5)	42 (87.5)
Comorbidities				NS
Luminal stenosis	31	4 (12.9)	27 (87.1)
No luminal stenosis	67	6 (9.0)	61 (91.0)
Severity				NS
Severe	42	3 (7.1)	39 (92.9)
Mild-moderate	56	7 (12.5)	49 (87.5)

NS: No significance. P268S+ Mutant P268S; P268S- Wild-type P268S.

^aP < 0.05 using Fisher's exact test.