Journal of Hematology, ISSN 1927-1212 print, 1927-1220 online, Open Access
Article copyright, the authors; Journal compilation copyright, J Hematol and Elmer Press Inc
Journal website http://www.thejh.org

Original Article

Volume 6, Number 4, October 2017, pages 73-80


The DAU Allele and Anti-D Alloimmunization Present With High Frequency in Brazilian Sickle Cell Disease Patients

Figure

Figure 1.
Figure 1. Distribution of 1025T>C, which is related to the DAR allele, and its association with a high risk of hospitalization.

Tables

Table 1. Synthetic Oligonucleotides Used to Amplify Exons 3, 5, 7 and 8 of the RHD and HBB Genes
 
GeneSequence (5′→3′)Exon RHD/mutation and HBB geneReference
F: forward; R: reverse. HBB: internal control beta globin gene.
RHDI4’F: TAAGCACTTCACAGAGCGGExon 5/667T>G[8]
EX5RR: TCTTGCTGATCTTCCCTTGG
RHDEX3FF: TATTCGGCTGGCCACCATGAExon 3/455A>C[8]
RHDI3RR: TTCTAATTTTGCCTGTAATC
RHD16FF: TCATAGTGTGGTCCGTAAACExon 7/1025T>C[9]
RHDEX7RR: CAATCATGCCCATTGCC
DAU 1BF: TTGGCCATCGTGATAGCTCACATExon 8/1136C>T[10]
DAUBR: GGAGATGGGGCACATAGACATC
HBB108F: CAAGGACAGGTACGGCTGTCGTCATCACPromoter region[11]
HBB109R: TTTTCCCAAGGTTTGAACTAGCTCTTCIVS-2 region

 

Table 2. RHD Gene Mutation Frequencies Among the SCD Patients and Healthy Individuals
 
GenotypeSCD (N = 193) (%)Healthy group (N = 220) (%)OR (95% CI); P
§Chi-squared Yates corrected. §§Fisher’s exact test. SCD: sickle cell disease; OR: odds ratio; CI: confidence interval.
667T>G
  T/T174 (90.1)211 (95.9)2.56 (1.07 - 6.29); 0.034§
  T/G15 (7.8)6 (2.7)
  G/D-4 (2.1)3 (1.4)
455A>C (DIIIa)
  A/A189 (97.9)220 (100)0.047§§
  A/C3 (1.5)0
  C/D-1 (0.6)0
1025T>C (DAR)
  T/T178 (92.2)211 (95.9)1.99 (0.8 - 5.05); 0.16§
  T/C12 (6.2)6 (2.7)
  C/D-3 (1.6)3 (1.4)
1136C>T (DAU)
  Absence150 (77.7)212 (96.4)7.65 (3.34 - 18.18); < 0.000§
  Presence43 (22.3)8 (3.6)
DIIIa/DAR or 1136C>T (DAU)
  Absence114 (78.6)**203 (92.3)3.25 (1.65 - 6.43); 0.002§
  Presence31 (21.4)**17 (7.7)

 

Table 3. Distributions of DIIIa, DAR and DAU Alleles Frequencies in the RHD Gene Among Individuals With HbSS and Healthy Individuals
 
GenotypesHbSS (N = 137) (%)Healthy individual group, N (%)OR (95% CI); P
**N = 92. §Chi-squared Yates corrected. §§Fisher’s exact test. OR: odds ratio; CI: confidence interval; HbSS: sickle cell anemia.
667T>G
  T/T122 (89.1)211 (95.9)2.88 (1.15 - 7.38); 0.021§
  T/G13 (9.5)6 (2.7)
  G/D-2 (1.4)3 ( 1.4 )
455A>C (DIIIa)
  A/A134 (97.8)220 (100)0.05§§
  A/C2 (1.4)0
  C/D-1 (0.8)0
1025T>C (DAR)
  T/T125 (91.2)211 (95.9)2.25 (0.86 - 5.95); 0.11§
  T/C11 (8.0)6 (2.7)
  C/D-1 (0.8)3 (1.4)
1136C>T (DAU)
  Absence109 (79.6)212 (96.4)6.81 (2.84 - 16.89); < 0.000§
  Presence28 (20.4)8 (3.6)
DIIIa/DAR or 1136C>T (DAU)
  Absence71 (77.2)**203 (92.3)3.53(1.67 - 7.48); < 0.000§
  Presence21 (22.8)**17 (7.7)

 

Table 4. Distributions of the DIIIa, DAR and DAU Alleles in SCD and Healthy Groups
 
RHD allelesSCD patients (n = 193)Healthy controls (n = 220)OR (95% CI)P
*Chi-squared Yates corrected. **Fisher’s exact test. OR: odds ratio; CI: confidence interval; SCD: sickle cell disease; HbSS: sickle cell anemia; HbSC: hemoglobin SC.
667T>G19 (9.8%)9 (4.1%)2.56 (1.07 - 6.29)0.034*
DIIIa4 (2.1%)0-0.047**
DAR15 (7.8%)9 (4.1%)1.99 (0.8 - 5.05)0.16*
1136C>T (DAU)43 (22.3%)8 (3.6%)7.65 (3.34 - 18.2)< 0.000*
HbSS (n = 137)
667T>G15 (10.9%)9 (4.1%)2.88 (1.15 - 7.38)0.021*
DIIIa3 (2.2%)0-0.05**
DAR12 (8.8%)9 (4.1%)2.25 (0.86 - 5.95)0.110*
1136C>T (DAU)28 (22.2%) (n = 126)8 (3.6%)6.81 (2.84 - 16.89)< 0.000*
HbSC (n = 56)
667T>G4 (7.1%)9 (4.1%)-0.31**
DIIIa1 (1.8%)0-0.20**
DAR3 (5.4%)9 (4.1%)-0.71**
1136C>T (DAU)14 (20.9%) (n = 67)8 (3.1%)7.00 (2.58 - 19.38)< 0.0001*

 

Table 5. Distribution of Partial D Genotype Frequencies in the SCD, HbSS and Healthy Groups
 
RHD gene allelesSCD (N = 193) (%)HbSS (N = 137) (%)Healthy individual group (N = 220) (%)
SCD: sickle cell disease; HbSS: sickle cell anemia.
DIIIa/D normal2 (1.0 )1 (0.7)-
DAR/D normal11 (5.7 )11 (8.0)6 (2.7)
DIIIa/D-1 (0.5)0-
DAR/D-3 (1.6)1 (0.7)3 (1.4)
DIIIa/DAU1 (0.5)1 (0.7)-
DAR/DAU1 (0.5)0-
DAU/ D-, D normal or DAU/DAU40 (20.7)27 (19.7)8 (3.6)

 

Table 6. Distribution of the “C”, “c”, “E”, and “e” Antigens of the Rh System and the DIIIa, DAR and DAU Alleles of the Gene RHD Among Sickle Cell Disease Patients
 
RHD allelescceeCceeccEeCcEeNot conclusive
DIIIa/D normal-1---
DAR/D normal522--
DIIIa/D-----1
DIIIa/DAU1----
DAR/DAU1----
DAU/D-, D normal or DAU/DAU14451-
Total (%)21 (56.8)7 (18.9)7 (18.9)1 (2.7)1 (2.7)