title: RHD genotyping and its implication in transfusion practice.
abstract: BACKGROUND
  The limitations of serology can be overcome by molecular typing. In order to evaluate the contribution of RH systematic genotyping and its implication in transfusion practice, a genotyping of D- blood donors was initiated.
  
  
  METHODS
  Blood samples were collected from 400 unrelated D- individuals. All samples were tested by RHD exon 10 PCR. In order to clarify the molecular mechanisms of RHD gene carrier, we applied molecular tools using different techniques: PCR-multiplex, and PCR-SSPs.
  
  
  RESULTS
  Among 400 D- subjects tested, 390 had RHD gene deletion; and 10 had RHD exon 10 of which seven were associated with the presence of the C or E antigens. Among D- carriers, we observed in five cases the presence of RHD-CE-Ds hybrid, in four cases the presence of pseudogene RHD ψ and in one case the presence of weak D type 4.
  
  
  CONCLUSIONS
  Since the majority of aberrant alleles were associated with C or E antigens and the preliminary infrastructure for molecular diagnostic were absent in all Tunisia territory, we recommend to reinforce transfusion practice to consider D- donors but C+/E+ antigens as D+ donors and the application of RHD molecular typing only to solve serologic problems.
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia.
authors.forename: Awatef
authors.initials: A
authors.lastname: Sassi
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia.
authors.forename: Mouna
authors.initials: M
authors.lastname: Ouchari
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia.
authors.forename: Batoul
authors.initials: B
authors.lastname: Houissa
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia.
authors.forename: Houda
authors.initials: H
authors.lastname: Romdhane
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia.
authors.forename: Saida
authors.initials: S
authors.lastname: Abdelkefi
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia.
authors.forename: Taher
authors.initials: T
authors.lastname: Chakroun
authors.affiliation: Centre Régional de Transfusion Sanguine, Hôpital Farhat Hached Sousse, Tunis, Tunisia. Electronic address: saloua.jemni@rns.tn.
authors.forename: Saloua
authors.initials: S
authors.lastname: Jemni Yacoub
mesh_terms: D001788:Blood Grouping and Crossmatching / Q000379:methods*; D001803:Blood Transfusion; D005091:Exons; D005260:Female; D060005:Genotyping Techniques; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012204:Rh-Hr Blood-Group System / Q000235:genetics*; D014416:Tunisia
chemical_list: D012204:Rh-Hr Blood-Group System; C026410:Rho(D) antigen
keywords: PCR-SSP; PCR-multiplex; RHD genotyping; Tunisia; Tunisian D negative blood donors