Treatment of pregnant women by RhD type

  • Janja Jerina Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia
  • Klara Železnik Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia
  • Tadeja Dovč Drnovšek Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia
  • Irena Bricl Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia
Keywords: variants D, serologic technicques, molecular-biologic methods (PCR-SSP), treatment algorithm

Abstract

An accurate determination of RhD (D) antigen (Ag) is important for pregnant women, transfusion recipients and blood donors. The decision to receive the pre- and postnatal prophylaxis with anti-D immunoglobulin (RhIG) is based on the D-type. Individuals can be classified as D-positive (D-pos) or D-negative (D-neg) based on the determination of Ag D but there are numerous other versions of Ag D as well (variants D, D-var). Allele variants of the RHD (D) gene can encode different forms of protein D. They are divided into weak, partial and Del. Ag D are determined on the basis of serological techniques. If in doubt, we use the molecular-biological methods for determining the D gene. The definitive definition of D-var is given on the basis of the molecular-biological methods.
From the peripheral venous blood samples taken from pregnant women, we roughly determined Ag D on plates (the Seraclone anti-D reagent) and on gel cards (the DiaClon ABO / D + Reverse Grouping test system (monoclonal antibodies) (Bio-Rad, Germany), with which we do not detect D category VI blood group). For 49 pregnant women we performed extended serological tests of D blood group (commercial ID-Partial RhD Typing Set (Bio-Rad, Germany)). We proceeded with molecular-biological methods: DNA isolation (utilisation of the BioRobot EZ1 and the commercial set EZ1 DNA Blood 350 μl Kit (Qiagen, Germany)), determination of the genotype D with the PCR-SSP method (commercial RBC-Ready Gene CDE, RBC -Ready Gene D weak and RBC-Ready Gene D AddOn (Inno-Train; Germany)), amplification (Veriti apparatus (Apllied BioSystems, USA)), product separation (electrophoretic system (BioRad, USA) on agarose gel (Sigma, Germany).
Our results showed that weak D forms represented 41 cases (83.7 %) and partial forms 8 cases (16.3 %) of all D-var. The most common D-var in pregnant women was weak D type 1 with 17 cases (34.7 %), followed by weak D type 3 with 13 cases (26.5 %) and weak D type 2 with 10 cases (20.4 %). The most common partial D form was D category VII with 5 cases (10.2 %).
Pregnant women and transfusion recipients who are carriers of weak D types 1, 2 or 3 can be safely treated as D-pos, while carriers of all other D-var can be treated as D-neg. Blood donors with D-var are treated as D-pos. By means of this algorithm, approximately 172 unnecessary RhIG applications can be prevented annually and adequate supplies of D-neg erythrocyte blood components can be maintained.

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Published
2020-01-09
How to Cite
1.
Jerina J, Železnik K, Dovč Drnovšek T, Bricl I. Treatment of pregnant women by RhD type. ZdravVestn [Internet]. 9Jan.2020 [cited 27Feb.2020];88(11-12):582-9. Available from: https://vestnik.szd.si/index.php/ZdravVest/article/view/2687
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Professional Article