RESUMO
BACKGROUND: Alloimmunisation and haemolytic transfusion reactions (HTRs) can occur in patients with sickle cell disease (SCD) despite providing phenotype-matched red blood cell (RBC) transfusions. Variant RBC antigen gene alleles/polymorphisms can lead to discrepancies in serological phenotyping. We evaluated differences between RBC antigen genotyping and phenotyping methods and retrospectively assessed if partial antigen expression may lead to increased risk of alloimmunisation and HTRs in SCD patients at a tertiary centre in Canada. METHODS: RBC antigen phenotyping and genotyping were performed by a reference laboratory on consenting SCD patients. Patient demographic, clinical and transfusion-related data were obtained from a local transfusion registry and chart review after research ethics board approval. RESULTS: A total of 106 SCD patients were enrolled, and 91% (n = 96) showed additional clinically relevant genotyping information when compared to serological phenotyping alone. FY*02N.01 (FY*B GATA-1) (n = 95; 90%) and RH variant alleles (n = 52, 49%; majority accompanied by FY*02N.01) were common, the latter with putative partial antigen expression in 25 patients. Variability in genotype-phenotype antigen prediction occurred mostly in the Rh system, notably with the e antigen (kappa: 0.17). Fifteen (14.2%) patients had a history of alloimmunisation, with five having HTR documented; no differences in clinical outcomes were found in patients with partial antigen expression. Genotype/extended-phenotype matching strategies may have prevented alloimmunisation events. CONCLUSION: We show a high frequency of variant alleles/polymorphisms in the SCD population, where genotyping may complement serological phenotyping. Genotyping SCD patients before transfusion may prevent alloimmunisation and HTRs, and knowledge of the FY*02N.01 variant allele increases feasibility of finding compatible blood.
Assuntos
Anemia Falciforme , Tipagem e Reações Cruzadas Sanguíneas , Sistema do Grupo Sanguíneo Duffy , Transfusão de Eritrócitos , Técnicas de Genotipagem , Receptores de Superfície Celular , Reação Transfusional , Adolescente , Adulto , Alelos , Anemia Falciforme/imunologia , Anemia Falciforme/terapia , Criança , Pré-Escolar , Sistema do Grupo Sanguíneo Duffy/genética , Sistema do Grupo Sanguíneo Duffy/imunologia , Feminino , Humanos , Masculino , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Estudos Retrospectivos , Fatores de Risco , Reação Transfusional/genética , Reação Transfusional/imunologia , Reação Transfusional/prevenção & controleAssuntos
Transfusão de Sangue Autóloga/métodos , Isoanticorpos/imunologia , Monócitos/imunologia , Vasculite Sistêmica/diagnóstico , Adolescente , Alelos , Doadores de Sangue , Incompatibilidade de Grupos Sanguíneos , Transfusão de Sangue Autóloga/efeitos adversos , Teste de Coombs/métodos , Procedimentos Cirúrgicos Eletivos/normas , Eritrócitos/metabolismo , Genótipo , Humanos , Masculino , Monócitos/metabolismo , Vasculite Sistêmica/cirurgia , Reação Transfusional/imunologia , Reação Transfusional/prevenção & controleRESUMO
BACKGROUND: RhIG prophylaxis for D- pregnant women prevents hemolytic disease of the newborn and typically depends on results of serologic D typing. Interpretation and follow-up of weak D serology is variable. Recent recommendations promote genotyping for RHD status determination in those with weak D serology. Canadian Blood Services performs comprehensive serologic prenatal testing in four provinces. Genotyping is used to determine D typing in patients with weak D. STUDY DESIGN AND METHODS: A serologic algorithm identified which patients require genotyping for RHD determination. Genotyping was performed on one of two commercially available platforms. RESULTS: Only 0.4% of D- patients met criteria for genotyping. Sixty-one percent were weak D Type 1, 2, or 3. Thirty percent had a partial or weak D other than Type 1, 2, or 3. Eleven had variants which remained unresolved. Seventeen were D+ and four were D-. CONCLUSIONS: Genotyping of patients with weak D serology led to an identified genotype in most patients. RhIG administration was avoided in 66% who were weak D Type 1, 2, or 3 or were D+. The use of a serologic algorithm to select patients for RHD genotyping identifies a majority of patients with weak D types not at risk for alloimmunization. This approach limits the number of genotyping investigations and the cost of providing classification for weak D types.
Assuntos
Tipagem e Reações Cruzadas Sanguíneas/classificação , Eritroblastose Fetal/prevenção & controle , Diagnóstico Pré-Natal/métodos , Sistema do Grupo Sanguíneo Rh-Hr/imunologia , Imunoglobulina rho(D)/análise , Adulto , Algoritmos , Tipagem e Reações Cruzadas Sanguíneas/métodos , Feminino , Genótipo , Humanos , Isoanticorpos/análise , Seleção de Pacientes , Gravidez , Adulto JovemRESUMO
BACKGROUND: The introduction of molecular methods into routine blood typing is prompting the identification of new blood group alleles. Discrepancies between the results of genotyping and serology or chance events uncovered during genotyping prompted additional investigations, which revealed six new RHCE variant alleles. STUDY DESIGN AND METHODS: Samples from eight blood donors, two patients (one prenatal), and a patient's relative, all of diverse racial origin, were analyzed by standard serology methods, targeted genotyping arrays, DNA sequencing, and allele-specific polymerase chain reaction. RESULTS: Six new RHCE alleles were identified, namely, RHCE*cE84A, RHCE*ce202G, RHCE*ce307T, RHCE*Ce377G, RHCE*ce697G,712G,733G,744C, and RHCE*Ce733G. CONCLUSION: While implementation of new assays in commercial genotyping platforms to detect the polymorphisms reported here may not be justified given their apparent rarity, software interpretative algorithms may benefit from the identification of new alleles for a more accurate determination of genotypes and prediction of phenotypes.
Assuntos
Alelos , População Negra/genética , Doadores de Sangue , Polimorfismo Genético , Sistema do Grupo Sanguíneo Rh-Hr/genética , População Branca/genética , Marcadores Genéticos , Genótipo , Técnicas de Genotipagem , Humanos , Fenótipo , Análise de Sequência de DNARESUMO
Correct donor D typing is critical to prevent recipient alloimmunization. No method can detect all variants, and the immunogenicity of many variants is unknown. Routine ABO and D serologic typings are performed in our laboratory by automated microplate testing. Until 2011, routine confirmation of D- status of first-time donors was performed by the manual tube indirect antiglobulin test (IAT); this was replaced by automated solid-phase testing including weak D testing by IAT. Selected donors are investigated by other methods. We describe four weak D type 67 (RHD*01W.67) donors whose samples tested as D- by automated microplate and manual methods but were later determined to be D+ by automated solid-phase and RHD gene analysis. Solid-phase serologic and molecular typing results of all four donors were identical. It was identified that the donors are of English-Irish descent; two are brothers and the others are cousins. Transfusion of blood from one of these donors likely resulted in alloimmunization to D in one of three recipients tested since no other documented exposures were identified. Lookback studies determined that two other D- recipients were not alloimmunized.
Assuntos
Doadores de Sangue , Sistema do Grupo Sanguíneo Rh-Hr/genética , Canadá , Teste de Histocompatibilidade , Humanos , MasculinoRESUMO
BACKGROUND: Variant alleles that do not produce RhCE antigens are rare. Consequently, they pose a challenge to transfusion when found in alloimmunized patients and make blood units valuable when found in donors. STUDY DESIGN AND METHODS: Five index cases and their relatives were studied by both serologic and molecular techniques. Genomic DNA was subjected to microarray genotyping, sequencing, exon scanning, and/or copy number determination assays to identify the RHCE allele(s) responsible for their D+ C- c- E- e- (D- -) phenotype. RESULTS: The five apparent D- - phenotypes were confirmed by molecular methods. Three of them contained unreported RHCE-null alleles, namely, RHCE*Ce-D(3-9)-Ce, RHCE*Ce87_93insT, and RHCE*cE221A. CONCLUSION: Molecular analysis of D- - phenotypes allows the identification of new RHCE-null variants. Conversely, detection of described RHCE-null variants facilitates confirmation of D- - phenotypes in patients and donors, helping improve transfusion safety.
Assuntos
Variação Genética , Sistema do Grupo Sanguíneo Rh-Hr/genética , Alelos , Feminino , Humanos , Recém-Nascido , Masculino , Linhagem , Fenótipo , GravidezAssuntos
Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Antígenos de Grupos Sanguíneos/genética , Códon sem Sentido , Mutação da Fase de Leitura , Proteínas de Neoplasias/genética , Alelos , Teste de Coombs , Éxons/genética , Feminino , Heterozigoto , Humanos , Masculino , Mutagênese Insercional , Paquistão/etnologia , Fenótipo , Gravidez , Análise de Sequência de DNA , Deleção de SequênciaAssuntos
Antígenos de Grupos Sanguíneos/genética , Tipagem e Reações Cruzadas Sanguíneas/métodos , DNA/isolamento & purificação , Eritrócitos/imunologia , Mucosa Bucal/química , Antígenos de Grupos Sanguíneos/biossíntese , DNA/genética , Humanos , Fenótipo , Polimorfismo de Nucleotídeo Único , Sensibilidade e Especificidade , Manejo de EspécimesRESUMO
BACKGROUND: AnWj is a high-prevalence red blood cell (RBC) antigen in the ISBT 901 series. Only nine reports of anti-AnWj have been published since it was first documented in 1972; two of these cases involved transfusion reactions. We present a case of a patient with aplastic anemia who developed anti-AnWj with clinically significant hemolysis after transfusion of AnWj-positive RBCs. STUDY DESIGN AND METHODS: A 56-year-old woman with aplastic anemia developed a pan-reactive antibody with clinically significant hemolysis after transfusion of RBCs. Investigations included antibody identification by serologic methods at the national immunohematology reference laboratory and the in vitro monocyte-monolayer assay (MMA) to predict clinical significance. RESULTS: The antibody was identified as having specificity for the AnWj antigen. The posttransfusion MMA indicated that the antibody had complement-fixing capability highly likely to cause clinically significant acute hemolytic transfusion reactions. The patient required long-term transfusion support with AnWj-negative RBCs and Lu(a-b-) RBCs of the In(Lu) type which have reduced AnWj expression. Cross-Canada and international collaboration was required to obtain rare units for transfusion. CONCLUSION: A case of acute hemolytic transfusion reaction caused by anti-AnWj is reported in a patient with aplastic anemia requiring allogeneic stem cell transplantation. This is the second documented case of anti-AnWj to cause a hemolytic transfusion reaction. The case demonstrates the complexity of managing patients with rare antibodies and the importance of international collaboration in the management of these difficult cases.