Introduction: Determination of RhD variants in blood donors, pregnant women, and newborns is important for transfusion strategies, in order to prevent RhD alloimmunisation and hemolytic disease of fetuses and newborns. Implementation of molecular RHD typing in two transfusion institutes is presented in this article, from Banja Luka (Bosnia and Herzegovina) and Belgrade (Serbia). Study Design and Methods: Blood donors’ RhD was checked by direct agglutination assays (tube) and indirect antiglobulin test (gel). Molecular RHD typing was performed by PCR-SSP with fluorometric signal detection in both centres. Donors were selected by weak RhD serological reactivity (Banja Luka, 85 samples; Belgrade, 62 samples) or serologically RhD-negative C/E-positive results (Banja Luka, 92 samples; Belgrade, 61 samples). Results: Among serologically determined weak D donors from the institute from Banja Luka, weak D type 3 was the most frequent (58.8%), followed by type 1 (35.3%) and DNB (1.2%), whereas results obtained at the Belgrade institute were distributed between weak D type 1 (41.9%), type 3 (30.7%), type 14 (6.5%), type 15 (1.6%), and DNB with anti-D (1.6%). In 17.7% of serologically typed weak D samples from the Belgrade institute, the molecular typing result was standard D. Additionally, RHD presence was detected in 9.8% of serologically RhD-negative, C/E-positive samples from both institutes. Conclusion: Rh molecular testing was successfully implemented in both blood transfusion institutes in Banja Luka and Belgrade. This study proved the efficiency of serological algorithms for weak D, as well as the presence of the RHD gene among serologically tested RhD-negative, C/E-positive samples.

1.
International Society of Blood Transfusion
. Table of blood group antigens v.8_180620,
2018
[accessed October 20, 2018], available from: http://www.isbtweb.org/fileadmin/user_upload/Red_Cell_Terminology_and_Immunogenetics/Table_of_blood_group_antigens_within_systems_v8_180620.pdf
2.
Okuda
H
,
Suganuma
H
,
Kamesaki
T
,
Kumada
M
,
Tsudo
N
,
Omi
T
, et al.
The analysis of nucleotide substitutions, gaps, and recombination events between RHD and RHCE genes through complete sequencing
.
Biochem Biophys Res Commun
.
2000
Aug
;
274
(
3
):
670
83
.
[PubMed]
0006-291X
3.
Daniels
G
.
Variants of RhD—current testing and clinical consequences
.
Br J Haematol
.
2013
May
;
161
(
4
):
461
70
.
[PubMed]
0007-1048
4.
Wagner
FF
,
Flegel
WA
.
RHD gene deletion occurred in the Rhesus box
.
Blood
.
2000
Jun
;
95
(
12
):
3662
8
.
[PubMed]
0006-4971
5.
Avent
ND
,
Ridgwell
K
,
Tanner
MJ
,
Anstee
DJ
.
cDNA cloning of a 30 kDa erythrocyte membrane protein associated with Rh (Rhesus)-blood-group-antigen expression
.
Biochem J
.
1990
Nov
;
271
(
3
):
821
5
.
[PubMed]
0264-6021
6.
Chérif-Zahar
B
,
Bloy
C
,
Le Van Kim
C
,
Blanchard
D
,
Bailly
P
,
Hermand
P
, et al.
Molecular cloning and protein structure of a human blood group Rh polypeptide
.
Proc Natl Acad Sci USA
.
1990
Aug
;
87
(
16
):
6243
7
.
[PubMed]
0027-8424
7.
Avent
ND
,
Reid
ME
.
The Rh blood group system: a review
.
Blood
.
2000
Jan
;
95
(
2
):
375
87
.
[PubMed]
0006-4971
8.
Daniels
G
.
Human Blood Groups
. 3rd ed.
Oxford
:
Wiley
;
2013
.
9.
Flegel
WA
.
Molecular genetics and clinical applications for RH
.
Transfus Apheresis Sci
.
2011
Feb
;
44
(
1
):
81
91
.
[PubMed]
1473-0502
10.
Reid
ME
,
Lomas-Francis
C
.
The Blood Group Antigen Facts Book
. 2nd ed.
San Diego
:
Academic Press
;
2003
.
11.
Denomme
GA
,
Dake
LR
,
Vilensky
D
,
Ramyar
L
,
Judd
WJ
.
Rh discrepancies caused by variable reactivity of partial and weak D types with different serologic techniques
.
Transfusion
.
2008
Mar
;
48
(
3
):
473
8
.
[PubMed]
0041-1132
12.
Judd
WJ
,
Moulds
M
,
Schlanser
G
.
Reactivity of FDA-approved anti-D reagents with partial D red blood cells
.
Immunohematology
.
2005
;
21
(
4
):
146
8
.
[PubMed]
0894-203X
13.
Jenkins
CM
,
Johnson
ST
,
Bellissimo
DB
,
Gottschall
JL
.
Incidence of weak D in blood donors typed as D positive by the Olympus PK 7200
.
Immunohematology
.
2005
;
21
(
4
):
152
4
.
[PubMed]
0894-203X
14.
Westhoff
CM
.
Review: the Rh blood group D antigen... dominant, diverse, and difficult
.
Immunohematology
.
2005
;
21
(
4
):
155
63
.
[PubMed]
0894-203X
15.
Credidio
DC
,
Pellegrino
J
 Jr
,
Castilho
L
.
Serologic and molecular characterization of D variants in Brazilians: impact for typing and transfusion strategy
.
Immunohematology
.
2011
;
27
(
1
):
6
11
.
[PubMed]
1930-3955
16.
Legler
TJ
,
Maas
JH
,
Köhler
M
,
Wagner
T
,
Daniels
GL
,
Perco
P
, et al.
RHD sequencing: a new tool for decision making on transfusion therapy and provision of Rh prophylaxis
.
Transfus Med
.
2001
Oct
;
11
(
5
):
383
8
.
[PubMed]
0958-7578
17.
Garratty
G
.
Do we need to be more concerned about weak D antigens?
Transfusion
.
2005
Oct
;
45
(
10
):
1547
51
.
[PubMed]
0041-1132
18.
Wagner
FF
,
Frohmajer
A
,
Ladewig
B
,
Eicher
NI
,
Lonicer
CB
,
Müller
TH
, et al.
Weak D alleles express distinct phenotypes
.
Blood
.
2000
Apr
;
95
(
8
):
2699
708
.
[PubMed]
0006-4971
19.
INTERNATIONAL FORUM
.
Testing for weak D
.
Vox Sang
.
2006
Feb
;
90
(
2
):
140
53
.
[PubMed]
0042-9007
20.
Gassner
C
,
Doescher
A
,
Drnovsek
TD
,
Rozman
P
,
Eicher
NI
,
Legler
TJ
, et al.
Presence of RHD in serologically D-, C/E+ individuals: a European multicenter study
.
Transfusion
.
2005
Apr
;
45
(
4
):
527
38
.
[PubMed]
0041-1132
21.
Judd
WJ
,
Johnson
ST
,
Storry
J
.
Judd’s methods in immunohematology
. 3rd ed.
Bethesda (MD)
:
AABB Press
;
2008
.
22.
Wagner
FF
,
Gassner
C
,
Müller
TH
,
Schönitzer
D
,
Schunter
F
,
Flegel
WA
.
Molecular basis of weak D phenotypes
.
Blood
.
1999
Jan
;
93
(
1
):
385
93
.
[PubMed]
0006-4971
23.
Araújo
F
,
Rodrigues
MJ
,
Monteiro
F
,
Chabert
T
,
Tavares
G
,
Sousa
G
, et al.
Weak D type 2 is the most prevalent weak D type in Portugal
.
Transfus Med
.
2006
Feb
;
16
(
1
):
63
7
.
[PubMed]
0958-7578
24.
St-Louis
M
,
Richard
M
,
Côté
M
,
Éthier
C
,
Long
A
.
Weak D type 42 cases found in individuals of European descent
.
Immunohematology
.
2011
;
27
(
1
):
20
4
.
[PubMed]
1930-3955
25.
Dogic
V
,
Bingulac-Popovic
J
,
Babic
I
,
Hundric-Haspl
Z
,
Jurakovic-Loncar
N
,
Mratinovic-Mikulandra
J
, et al.
Distribution of weak D types in the Croatian population
.
Transfus Med
.
2011
Aug
;
21
(
4
):
278
9
.
[PubMed]
0958-7578
26.
Johnson
ST
. Partial D & Weak D Picking Up the Rhesus Pieces. Heart of America Association of Blood Banks, April 24,
2012
27.
Sandler
SG
,
Flegel
WA
,
Westhoff
CM
,
Denomme
GA
,
Delaney
M
,
Keller
MA
, et al.;
College of American Pathologists Transfusion Medicine Resource Committee Work Group
.
It’s time to phase in RHD genotyping for patients with a serologic weak D phenotype
.
Transfusion
.
2015
Mar
;
55
(
3
):
680
9
.
[PubMed]
0041-1132
28.
Agre
PC
,
Davies
DM
,
Issitt
PD
,
Lamy
BM
,
Schmidt
PJ
,
Treacy
M
, et al.
A proposal to standardize terminology for weak D antigen
.
Transfusion
.
1992
Jan
;
32
(
1
):
86
7
.
[PubMed]
0041-1132
29.
Fung
MK
,
Grossman
BJ
,
Hillyer
CD
,
Westhoff
CM
, editors
.
Technical Manual 18
.
Bethesda (MD)
:
AABB
;
2014
.
30.
Mota
M
,
Fonseca
NL
,
Rodrigues
A
,
Kutner
JM
,
Castilho
L
.
Anti-D alloimmunization by weak D type 1 red blood cells with a very low antigen density
.
Vox Sang
.
2005
Feb
;
88
(
2
):
130
5
.
[PubMed]
0042-9007
31.
Milkins
C
,
Berryman
J
,
Cantwell
C
,
Elliott
C
,
Haggas
R
,
Jones
J
, et al.;
British Committee for Standards in Haematology
.
Guidelines for pre-transfusion compatibility procedures in blood transfusion laboratories
.
Transfus Med
.
2013
Feb
;
23
(
1
):
3
35
.
[PubMed]
0958-7578
32.
de Vries
R
,
Haas
F
;
working group for revision of the Dutch Blood Transfusion Guideline 2011
.
English translation of the dutch blood transfusion guideline 2011
.
Clin Chem
.
2012
Aug
;
58
(
8
):
1266
7
.
[PubMed]
0009-9147
33.
Wagner
FF
,
Eicher
NI
,
Jørgensen
JR
,
Lonicer
CB
,
Flegel
WA
.
DNB: a partial D with anti-D frequent in Central Europe
.
Blood
.
2002
Sep
;
100
(
6
):
2253
6
.
[PubMed]
0006-4971
34.
Opoku-Okrah
C
,
Amidu
N
,
Amoah-Sakyi
S
.
Detection of weak D (Du) phenotype among Rh-D negative males and females in Kumasi, Ghana
.
Journal of Science & Technology (Ghana)
.
2008
;
28
:
34
40
.
35.
Hussein
E
,
Teruya
J
.
Weak D types in the Egyptian population
.
Am J Clin Pathol
.
2013
Jun
;
139
(
6
):
806
11
.
[PubMed]
0002-9173
36.
El Wafi
M
,
El Housse
H
,
Nourichafi
N
,
Bouisk
K
,
Benajiba
M
,
Habti
N
.
Prevalence of weak D phenotype among D negative C/E+ blood donors in Morocco
.
Int J Blood Transfus Immunohematol
.
2016
;
6
:
3
7
.
37.
Wagner
FF
,
Flegel
WA
.
The Rhesus Site.
Transfus Med Hemother
2014
;41(5): 357-363. PMID: 25538538, URL: The Human RhesusBase, version 2.3, http://www.rhesusbase.info/, update 2018-05-30
38.
Sandler
SG
,
Chen
LN
,
Flegel
WA
.
Serological weak D phenotypes: a review and guidance for interpreting the RhD blood type using the RHD genotype
.
Br J Haematol
.
2017
Oct
;
179
(
1
):
10
9
.
[PubMed]
0007-1048
39.
Ouchari
M
,
Srivastava
K
,
Romdhane
H
,
Jemni Yacoub
S
,
Flegel
WA
.
Transfusion strategy for weak D Type 4.0 based on RHD alleles and RH haplotypes in Tunisia
.
Transfusion
.
2018
Feb
;
58
(
2
):
306
12
.
[PubMed]
0041-1132
40.
Crottet
SL
,
Henny
C
,
Meyer
S
,
Still
F
,
Stolz
M
,
Gottschalk
J
, et al.
Implementation of a mandatory donor RHD screening in Switzerland
.
Transfus Apheresis Sci
.
2014
Apr
;
50
(
2
):
169
74
.
[PubMed]
1473-0502
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