Introduction
Materials and methods
Study subjects
DNA extraction and genotyping
Locus
|
Primer
|
Annealing temperature
|
Method
|
Product size(bp)
|
---|---|---|---|---|
+874 T/A | T allele primer: 5′-TTCTTACAACACAAATACAATACT-3′ | 61°C,56°C | SSP-PCR | AA,AT,TT:303 |
(rs2430561) | A allele primer: 5′-TTCTTACAACACAAATACAATACA-3′ | |||
common primer: 5′-TCCCATATAAACCTATAATATGCA-3′ | ||||
inner reference F: 5′-CTTCAACACCCCAGCCTAGTAC-3′ | Inner reference: 400 | |||
inner reference R: 5′-CCTCAGGGCAGCGAGACC-3′ | ||||
+2109A/G (rs1861494) | F: 5′-GAGAGGTAAGGTGAGAGAGAACC-3′ | 55°C | RFLP-PCR (MspI) | AA:267 |
R: 5′-AGCAAAGAAGGTCTACAAACT-3′ | AG:267 + 245 + 22 | |||
GG:245 + 22 |
Statistical analysis
Results
Characteristics of the study subjects
Factors
|
Control
|
CHB
|
LC
|
P
value
|
---|---|---|---|---|
N = 173
|
N = 129
|
N = 126
| ||
Age(mean ± SD)*
| 37.78 ± 11.71 | 37.71 ± 11.55 | 46.55 ± 10.28 | 0.000 |
Gender**
| ||||
Male | 149 (86.1%) | 115 (89.1%) | 106 (86.4) | 0.497 |
Femle | 24 (13.9%) | 14 (10.9%) | 20 (13.6) | |
Smoking**
| ||||
No | 127 (73.4%) | 96 (74.4) | 101 (80.2) | 0.373 |
Yes | 46 (26.6%) | 33 (25.6) | 25 (19.8) | |
Alcohol**
| ||||
No | 121 (69.9%) | 88 (68.2) | 91 (70.1) | 0.782 |
Yes | 52 (30.1%) | 41 (31.8) | 35 (29.9) |
Genotype and allele frequency distributions of the IFN-γ polymorphisms
Polymorphisms
|
Frequencyn (%)
|
CHB vs. Control
|
HBV-LC vs. Control
| |||||
---|---|---|---|---|---|---|---|---|
LC = 126
|
CHB = 129
|
Control = 173
|
P
|
OR (95% CI)
|
P
|
OR (95% CI)
| ||
+874(T/A)
| ||||||||
Genotypes | ||||||||
AA | 90 (71.4) | 84 (65.1) | 116 (67.1) | 1Ref
| 1Ref
| |||
AT | 32 (25.4) | 36 (27.9) | 47 (27.2) | 0.781 | 1.086 (0.607-1.943) | 0.588 | 0.863 (0.507-1.470) | |
TT | 4 (3.2) | 9 (7.0) | 10 (5.8) | 0.481 | 1.458 (0.511-4.159) | 0.264 | 0.505 (0.152-1.675) | |
Allele | ||||||||
A | 212 (84.1) | 204 (79.1) | 279 (80.6) | 1Ref
| 1Ref
| |||
T | 40 (15.9) | 54 (20.9) | 67 (19.4) | 0.480 | 1.176 (0.750-1.844) | 0.250 | 0.775 (0.505-1.196) | |
Dominant model AT + TT vs AA | 0.617 | 1.148 (0.668-1.974) | 0.389 | 0.801 (0.483-1.328) | ||||
Recessive model TT vs AA + AT | 0.505 | 1.421 (0.506-3.991) | 0.291 | 0.526 (0.160-1.730) | ||||
+2109 (A/G)
| ||||||||
Genotypes | ||||||||
AA | 79 (62.7) | 72 (55.8) | 82 (47.4) | 1Ref
| 1Ref
| |||
AG | 40 (32.7) | 43 (33.3) | 68 (39.3) | 0.640 | 0.874 (0.498-1.534) | 0.071 | 0.630 (0.381-1.041) | |
GG | 7 (5.6) | 14 (10.9) | 23 (13.3) | 0.256 | 0.625 (0.278-1.407) | 0.014 | 0.321 (0.130-0.793) | |
Allele | ||||||||
A | 198 (78.6) | 187 (72.5) | 232 (67.6) | 1Ref
| 1Ref
| |||
G | 54 (21.4) | 71 (27.5) | 111 (32.4) | 0.232 | 0.785 (0.528-1.167) | 0.003 | 0.565 (0.388-0.825) | |
Dominant model AG + GG vs AA | 0.392 | 0.799 (0.478-1.336) | 0.013 | 0.551 (0.344-0.883) | ||||
Recessive model GG vs AA + AG | 0.295 | 0.659 (0.301-1.440) | 0.034 | 0.385 (0.159-0.930) |
Association between the IFN-γ polymorphisms and HBV-related diseases
Haplotype analyses
Haplotype
|
CHB vs control
|
LC vs control
| ||||||
---|---|---|---|---|---|---|---|---|
Case (%)
|
Control (%)
|
P
|
OR [95% CI]
|
Case (%)
|
Control (%)
|
P
|
OR [95% CI]
| |
A+874A+2109
| 148 (57.4) | 186 (53.8) | 0.384 | 1.155 [0.835 ~ 1.599] | 160 (63.4) | 186 (53.8) | 0.019 | 1.485 [1.065 ~ 2.070] |
A+874G+2109
| 56 (21.7) | 93 (26.8) | 0.148 | 0.756 [0.517 ~ 1.105] | 52 (20.7) | 92 (26.8) | 0.087 | 0.714 [0.485 ~ 1.051] |
T+874A+2109
| 39 (15.1) | 46 (13.2) | 0.512 | 1.166 [0.735 ~ 1.850] | 38 (15.2) | 45 (13.2) | 0.495 | 1.175 [0.739 ~ 1.868] |
T+874G+2109
| 15 (5.8) | 21 (6.1) | 0.876 | 0.948 [0.480 ~ 1.872] | 2 (0.7) | 21 (6.1) | 0.001 | 0.106 [0.022 ~ 0.502] |
Genotype and allele frequencies among different races
SNPs
|
Number
|
Genotype frequency
|
P
|
Allele frequency
|
P
| |||
---|---|---|---|---|---|---|---|---|
+874 T/A
|
AA
|
AT
|
TT
|
A
|
T
| |||
This study | 173 | 116 (67.1) | 47 (27.2) | 10 (5.8) | 1Ref
| 279 (80.6) | 67 (19.4) | 1Ref
|
Korea [20] | 201 | 151 (75.1) | 47 (23.4) | 3 (1.5) | 0.043 | 249 (82.5) | 53 (17.5) | 0.553 |
Brazil [18] | 202 | 79 (39.1) | 82 (40.6) | 41 (20.3) | 0.000 | 240 (59.4) | 164 (40.6) | 0.000 |
India [15] | 146 | 52 (35.6) | 77 (52.7) | 17 (11.6) | 0.000 | 181 (62.0) | 111 (38.0) | 0.032 |
Tunisia [19] | 103 | 33 (32.0) | 47 (45.6) | 23 (22.3) | 0.000 | 113 (54.9) | 93 (45.1) | 0.000 |
+2109A/G
|
AA
|
AG
|
GG
|
A
|
G
| |||
This study | 173 | 112 (64.7) | 52 (30.1) | 9 (5.2) | 1Ref
| 232 (67.6) | 111 (32.4) | 1Ref
|
HCB | 45 | 21 (46.7) | 18 (40.0) | 6 (13.3) | 0.040 | 60 (66.7) | 30 (33.3) | 0.008 |
JPT | 44 | 16 (36.4) | 13 (29.5) | 15 (34.1) | 0.000 | 45 (51.1) | 43 (48.9) | 0.000 |
CEU | 58 | 23 (39.7) | 31 (53.4) | 4 (6.9) | 0.003 | 77 (66.4) | 39 (33.6) | 0.003 |
YRI | 59 | 47 (79.7) | 12 (20.3) | 0 (0) | 0.050 | 106 (89.8) | 12 (10.2) | 0.013 |