Background
Materials and methods
Study site
Study population
Sample size determination and data collection
Determination of infant EBV infection status
Laboratory procedures
Sickle-cell polymerase chain reaction genotyping test
Alpha-thalassemia genotyping
Multiplex PCR amplification for glucose-6-Phosphate dehydrogenase gene mutation
Mutations | Substitutions | Primer sequence (5’- 3’) | Size (bp) | |
---|---|---|---|---|
Group A1
| Viangchan Chinese | 871 G > A 1024 C > T | CCAACTCAACACCCAAGGAGC GGCATGCCCAGTTCTGCCTTG |
280
|
Group A2
| Mahidol Coimbra | 487 G > A 592 C > T | TGAATGATGCAGCTCTGATCC CCAGGTGAGGCTCCTGAGTAC |
293
|
Group B
| Union Kaiping | 1360 C > T 1388 G > A | TGGCATCAGCAAGACACTCTC GGAGAGGCATGAGGTAGCTCC |
384
|
G6PD african A- genotyping assay
MSP-2 genotyping assay
Data analysis
Analyses of sickle cell, G6PD, α-thalassemia genotypes, and EBV acquisition
Results
Demographic and clinical characteristics of study participants
Maternal characteristics | Summary | ||
---|---|---|---|
Median age in years (± S. D) at enrollment | 22.0 | ||
Education
| |||
Primary | 25.9% (21) | ||
Secondary | 53.1% (43) | ||
Polytechnic | 14.8% (12) | ||
College and higher | 6.2% (5) | ||
Gestation age at enrolment
| |||
1st trimester (< 14 weeks) | 12% (10) | ||
2nd trimester (14–27 weeks) | 65.4% (53) | ||
3rd trimester (≥ 28 weeks) | 22.2% (18) | ||
Marital status
| |||
Single | 29.6% (24) | ||
Married | 70.4% (57) | ||
Pathogenic protozoan and helminth infections
| |||
Trichomonas vaginalis
| 2.5% (2) | ||
Ascaris lumbricoides
| 3.7% (3) | ||
Trichuris trichiura
| 1.2% (1) | ||
Ascaris lumbricoides
| 3.7% (3) | ||
Trichuris trichiura
| 1.2% (1) | ||
Schistosoma mansoni
| 1% (2) | ||
Infant characteristics
|
Summary
| ||
Gender | Male n = 41(%) | Female n = 40 (%) | |
EBV Positive at 12 months | 46.7% (14) | 53.3% (16) | |
EBV Negative at 12 months | 52.9% (27) | 47.1% (24) | |
In utero malaria exposed | 38.1% (8) | 61.9% (13) | |
Malaria non exposed | 55.0% (33) | 45.0% (27) | |
P. falciparum infected at 12months | 38.1% (8) | 61.9% (13) | |
Alpha thalassemia (-α3.7/αα) | 50.0% (17) | 48.6% (17) | |
Hb AS | 41.2% (7) | 58.8% (10) | |
G6PD A (376T > A) | 50.9% (28) | 49.1% (27) | |
G6PD A- (202 C > T/376T > A) | 36.8% (7) | 63.2% (12) |
Distribution of alpha thalassemia, SCT, G6PD, and MSP-2 mutations based on EBV status < 6 months of age
EBV status < 6 months | EBV status ≥ 6-12months | ||||||
---|---|---|---|---|---|---|---|
Gene mutation type | n | EBV negative n (%) | EBV Positive n (%) | P-value | EBV negative n (%) | EBV Positive n (%) | p-value |
Thalassemia
| |||||||
αα/αα | 46 | 9 (19.6) | 37 (80.4) | b0.615 | 27 (58.7) | 19 (41.3) | a0.413 |
-α3.7/αα | 34 | 4 (11.8) | 30 (88.2) | 24 (70.6) | 10 (29.4) | ||
Sickle cell Status
| |||||||
Hb AA | 62 | 10 (16.1) | 52 (83.9) | b1.000 | 37 (74.0) | 25 (40.3) | b0.249 |
Hb AS | 17 | 3 (17.6) | 14 (82.4) | 13 (26.0) | 4 (13.8) | ||
G6PD Group A1
| |||||||
Viangchan 871G > A /Chinese 1024 C > T -Negative | 9 | 3(30) | 6 (66.7) | b0.153 | 5 (6.2) | 4 (4.9) | b0.616 |
Viangchan 871G > A /Chinese 1024 C > T -Positive | 72 | 10(14.1) | 62 (86.1) | 46 (56.8) | 26 (32.1) | ||
G6PD group A2
| |||||||
Mahidol 487G > A/Coimbra 592 C > T -Negative | 5 | 0 (0) | 5 (7.4) | b0.407 | 3 (3.7) | 2 (2.5) | b0.442 |
Mahidol 487G > A/Coimbra 592 C > T -Positive | 76 | 13 (16.9) | 63 (82.9) | 48 (59.3) | 28 (34.6) | ||
G6PD group B
| |||||||
Union 1360 C > T/Kaiping 1388G > A)-Negative | 15 | 1(6.7) | 14 (93.3) | b0.197 | 5 (6.2) | 10 (12.3) | a0.494 |
Union 1360 C > T /Kaiping 1388G > A)-Positive | 66 | 12(18.2) | 54 (81.8) | 41 (50.6) | 25 (30.9) | ||
G6PD African A-
| |||||||
A- | 19 | 6 (31.6) | 13 (68.4) | a0.084 | 14 (17.5) | 5 (6.3) | a0.280 |
A | 55 | 7(12.7) | 48 (87.3) | 31 (38.8) | 24 (30.0) | ||
B | 6 | 0 (0.0) | 6 (100) | 5 (6.3) | 1 (1.3) | ||
MSP-2
| |||||||
FC27 Negative | 50 | 7 (14) | 43 (86) | b0.914 | 31 (59.6) | 19 (65.5) | a0.814 |
FC27 Positive | 20 | 3 (15) | 17 (85) | 13 (25.0) | 7 (24.1) | ||
3D7 Negative | 29 | 4 (13.8) | 25 (86.2) | b0.921 | 16 (30.8) | 13 (44.8) | a0.263 |
3D7 Positive | 41 | 6 (14.6) | 35 (85.4) | 28 (68.3) | 13 (31.7) |
Distribution of alpha thalassemia, SCT, G6PD and MSP-2 mutations based on EBV status ≥ 6-12months of age
Association between carriage of hemoglobinopathy gene, MSP-2 allele, and EBV acquisition < 6 months of age
EBV status < 6 months | EBV status ≥ 6-12months | ||||||
---|---|---|---|---|---|---|---|
Gene mutation type | n | OR | 95% CI | P-value | OR | 95% CI | P-value |
α- thalassemia
| |||||||
αα/αα | 46 |
Ref
| - | - |
Ref
| - | - |
-α3.7/αα | 34 | 1.824 | 0.511–6.512 | 0.354 | 0.681 | 0.256–1.812 | 0.442 |
Sickle cell Status
| |||||||
Hb AA | 62 |
Ref
| - | - |
Ref
|
-
|
-
|
Hb AS | 17 | 0.897 | 0.217–3.708 | 0.881 | 0.513 | 0.143–1.836 | 0.305 |
G6PD Group A1
| |||||||
871G > A/1024C > T-Negative | 10 |
Ref
| - | - |
Ref
| - | - |
871G > A/1024C > T-Positive | 71 | 2.614 | 0.578–11.820 | 0.212 | 0.640 | 0.079–5.205 | 0.677 |
G6PD group A2
| |||||||
487G > A/592C > T-Negative | 4 |
Ref
| - | - |
Ref
| - | - |
487G > A/592C > T-Positive | 77 | 0.000 | 0.000 | 0.999 | 0.948 | 0.240–3.755 | 0.940 |
G6PD group B
| |||||||
1360C > T /1388G > A-Negative | 15 |
Ref
| - | - |
Ref
| - | - |
1360C > T /1388G > A-Positive | 66 | 0.321 | 0.038–2.686 | 0.295 | 1.221 | 0.325–4.592 | 0.768 |
G6PD African A-
| |||||||
B | 6 |
Ref
|
-
|
-
|
Ref
|
-
|
-
|
A- | 19 | xx | xx | 1.000 | 0.000 | 0.000 | 0.235 |
A | 55 | xx | xx | 0.999 | 0.278 | 0.030–2.544 | 0.257 |
B and A | 61 |
Ref
| - | - |
Ref
| ||
A- | 19 | 0.281 | 0.081–0.978 |
0.046
| 0.223 | 0.153–1.549 | 0.487 |
MSP-2
| |||||||
FC27 Negative | 50 |
Ref
|
-
|
-
|
Ref
|
-
|
-
|
FC27 Positive | 20 | 0.922 | 0.213–3.990 | 0.914 | 0.780 | 0.256–2.373 | 0.662 |
3D7 Negative | 29 |
Ref
|
-
|
-
|
Ref
|
-
|
-
|
3D7 Positive | 41 | 0.933 | 0.238–3.656 | 0.921 | 0.549 | 0.201–1.496 | 0.241 |