Assuntos
Sistema ABO de Grupos Sanguíneos/sangue , Incompatibilidade de Grupos Sanguíneos/sangue , Hemólise , Transfusão de Plaquetas/efeitos adversos , Reação Transfusional/sangue , Incompatibilidade de Grupos Sanguíneos/terapia , Ponte de Artéria Coronária , Humanos , Masculino , Pessoa de Meia-Idade , Reação Transfusional/terapiaRESUMO
BACKGROUND AND OBJECTIVES: Variant RHD genes associated with the weak D phenotype can result in complete or partial D-epitope expression on the red cell. This study examines the genetic classification in Australian blood donors with a weak D phenotype and correlates RHD variants associated with the weak D phenotype against D-epitope profile. MATERIALS AND METHODS: Following automated and manual serology, blood samples from donors reported as 'weak D' (n = 100) were RHD genotyped by a commercial SNP-typing platform and Sanger sequencing. Two commercial anti-D antibody kits were used for extended serological testing for D-epitope profiles. RESULTS: Three samples had wild-type RHD exonic sequences, and 97 samples had RHD variants. RHD*weak D type 1, RHD*weak D type 2 or RHD*weak D type 3 was detected in 75 donors. The remaining 22 samples exhibited 17 different RHD variants. One donor exhibited a novel RHD*c.939+3A>C lacking one D-epitope. Weak D types 1·1, 5, 15, 17 and 90 showed a partial D-epitope profile. CONCLUSION: The array of RHD variants detected in this study indicated diversity in the Australian donor population that needs to be accommodated for in future genotyping strategies.
Assuntos
Doadores de Sangue/estatística & dados numéricos , Sistema do Grupo Sanguíneo Rh-Hr/genética , Alelos , Austrália , Sequência de Bases , Transfusão de Sangue , DNA/química , DNA/isolamento & purificação , DNA/metabolismo , Epitopos/imunologia , Epitopos/metabolismo , Éxons , Frequência do Gene , Genótipo , Humanos , Isoanticorpos/sangue , Fenótipo , Polimorfismo de Nucleotídeo Único , Imunoglobulina rho(D)/sangue , Análise de Sequência de DNA , Testes SorológicosRESUMO
BACKGROUND AND OBJECTIVES: Duffy blood group phenotypes can be predicted by genotyping for single nucleotide polymorphisms (SNPs) responsible for the Fy(a) /Fy(b) polymorphism, for weak Fy(b) antigen, and for the red cell null Fy(a-b-) phenotype. This study correlates Duffy phenotype predictions with serotyping to assess the most reliable procedure for typing. MATERIALS AND METHODS: Samples, n = 155 (135 donors and 20 patients), were genotyped by high-resolution melt PCR and by microarray. Samples were in three serology groups: 1) Duffy patterns expected n = 79, 2) weak and equivocal Fy(b) patterns n = 29 and 3) Fy(a-b-) n = 47 (one with anti-Fy3 antibody). RESULTS: Discrepancies were observed for five samples. For two, SNP genotyping predicted weak Fy(b) expression discrepant with Fy(b-) (Group 1 and 3). For three, SNP genotyping predicted Fy(a) , discrepant with Fy(a-b-) (Group 3). DNA sequencing identified silencing mutations in these FY*A alleles. One was a novel FY*A 719delG. One, the sample with the anti-Fy3, was homozygous for a 14-bp deletion (FY*01N.02); a true null. CONCLUSION: Both the high-resolution melting analysis and SNP microarray assays were concordant and showed genotyping, as well as phenotyping, is essential to ensure 100% accuracy for Duffy blood group assignments. Sequencing is important to resolve phenotype/genotype conflicts which here identified alleles, one novel, that carry silencing mutations. The risk of alloimmunisation may be dependent on this zygosity status.
Assuntos
Algoritmos , Sistema do Grupo Sanguíneo Duffy/genética , Receptores de Superfície Celular/genética , Alelos , Sequência de Bases , Estudos de Associação Genética , Humanos , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Transição de Fase , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNARESUMO
BACKGROUND AND OBJECTIVES: An Australian Caucasian blood donor consistently presented a serology profile for the Duffy blood group as Fy(a+b+) with Fy(a) antigen expression weaker than other examples of Fy(a+b+) red cells. Molecular typing studies were performed to investigate the reason for the observed serology profile. MATERIAL AND METHODS: Blood group genotyping was performed using a commercial SNP microarray platform. Sanger sequencing was performed using primer sets to amplify across exons 1 and 2 of the FY gene and using allele-specific primers. RESULTS: The propositus was genotyped as FY*A/B, FY*X heterozygote that predicted the Fy(a+b+(w) ) phenotype. Sequencing identified the 265T and 298A variants on the FY*A allele. This link between FY*A allele and 265T was confirmed by allele-specific PCR. CONCLUSION: The reduced Fy(a) antigen reactivity is attributed to a FY*A allele-carrying 265T and 298A variants previously defined in combination only with the FY*B allele and associated with weak Fy(b) antigen expression. This novel allele should be considered in genotyping interpretative algorithms for generating a predicted phenotype.
Assuntos
Doadores de Sangue , Sistema do Grupo Sanguíneo Duffy/genética , Polimorfismo de Nucleotídeo Único , Algoritmos , Alelos , Austrália , Genótipo , Técnicas de Genotipagem/métodos , Humanos , Dados de Sequência Molecular , Fenótipo , População Branca/genéticaRESUMO
OBJECTIVE: To optimize the delay time before the initiation of arterial phase scan in the detection of focal liver lesions in contrast enhanced 5 phase liver CT using the bolus tracking technique. PATIENTS AND METHODS: Delay - the interval between threshold enhancement of 100 hounsfield unit (HU) in the abdominal aorta and commencement of the first arterial phase scan. Using a 16 slice CT scanner, a plain CT of the liver was done followed by an intravenous bolus of 120 ml nonionic iodinated contrast media (370 mg I/ml) at the rate of 4 mL/s. The second phase scan started immediately after the first phase scan. The portal venous and delay phases were obtained at a fixed delay of 60 s and 90 s from the beginning of contrast injection. Contrast enhancement index (CEI) and subjective visual conspicuity scores for each lesion were compared among the three groups. RESULTS: 84 lesions (11 hepatocellular carcinomas, 17 hemangiomas, 39 other hypervascular lesions and 45 cysts) were evaluated. CEI for hepatocellular carcinomas appears to be higher during the first arterial phase in the 6 seconds delay group. No significant difference in CEI and mean conspicuity scores among the three groups for hemangioma, other hypervascular lesions and cysts. CONCLUSION: The conspicuity of hepatocellular carcinomas appeared better during the early arterial phase using a bolus tracking technique with a scan delay of 6 seconds from the 100 HU threshold in the abdominal aorta.
RESUMO
Objective: To optimize the delay time before the initiation of arterial phase scan in the detection of focal liver lesions in contrast enhanced 5 phase liver CT using the bolus tracking technique. Patients and Methods: Delay - the interval between threshold enhancement of 100 hounsfield unit (HU) in the abdominal aorta and commencement of the first arterial phase scan. Using a 16 slice CT scanner, a plain CT of the liver was done followed by an intravenous bolus of 120 ml nonionic iodinated contrast media (370 mg I/ml) at the rate of 4 mL/s. The second phase scan started immediately after the first phase scan. The portal venous and delay phases were obtained at a fixed delay of 60 s and 90 s from the beginning of contrast injection. Contrast enhancement index (CEI) and subjective visual conspicuity scores for each lesion were compared among the three groups. Results: 84 lesions (11 hepatocellular carcinomas, 17 hemangiomas, 39 other hypervascular lesions and 45 cysts) were evaluated. CEI for hepatocellular carcinomas appears to be higher during the first arterial phase in the 6 seconds delay group. No significant difference in CEI and mean conspicuity scores among the three groups for hemangioma, other hypervascular lesions and cysts. Conclusion: The conspicuity of hepatocellular carcinomas appeared better during the early arterial phase using a bolus tracking technique with a scan delay of 6 seconds from the 100 HU threshold in the abdominal aorta.