The study was aimed to analyze Del phenotype of RhD (-) unrelated blood donors. RhD (-) was initially screened by routine serological test and confirmed by indirect antiglobulin test (IAT). Del phenotype was detected by hot-ether absorption-elution test. The results indicated that 106 RhD (-) samples were confirmed out of 38526 donors, and 28 cases were Del detected by hot-ether absorption-elution test. The incidence of Del in RhD (-) samples was 26.41%, The serological phenotypes of Del were Ccee (78.57%), CCee (14.29%) and CcEe (7.14%) respectively. In conclusion, the detection of Del by using hot-ether absorption-elution test is very important for reasonable application of RhD (-) blood. There is difference in Del phenotypes of populations in different regions of China and Japan.
Asian Continental Ancestry Group ; Blood Donors ; Humans ; Phenotype ; Rh-Hr Blood-Group System ; genetics ; immunology

To investigate the molecular basis of partial D phenotypes in Chinese, D variants with weak D expression was screened by using indirect anti-human globulin test (IAT) method, the polymerase chain reaction-sequence specific primer (PCR-SSP) method was employed to amplify RHD specific exons and their flanking regions. The amplification products were sequenced directly to determine the molecular basis of D variants. The results showed that ten cases of partial D phenotypes, including one case of D Va (Kou.), one case of D Va (Hus.), one case of D Va-like (YH.), and seven cases of D VI type III, were detected from 22 cases of weak D phenotype respectively. All ten cases of partial D phenotypes had one RHD allele deleted. In conclusion, the molecular basis of ten cases of partial D phenotype was confirmed, including D Va (Kou.) and D Va-like (YH.) phenotypes reported firstly in Chinese population.
Alleles ; Asian Continental Ancestry Group ; Base Sequence ; Humans ; Molecular Sequence Data ; Mutation ; Phenotype ; Rh-Hr Blood-Group System ; genetics ; immunology

In order to analyze the genotype of RhD-negative blood donors with immune antibodies in Harbin, the voluntary blood donors from 1 April 2008 to 30 september 2011 were detected serologically to determine the RhD-negative donors. The blood donors confirmed to be RhD negative were detected to screen the immune antibodies, the samples with immune antibodies were analyzed by PCR-SSP and DNA sequencing to detect RhD genotype. The results showed that the 12 cases of the immune antibodies (0.95%) were screened out from 1265 cases of RhD-negative donors, among which 9 cases showed anti-D-antibody, 3 cases showed anti-(D+C) antibody; 10 cases were RhD-negative, 2 cases were RHD 711D(el)C. It is concluded that RhD negative and RHD 711D(el)C are easy to be immunized to produce the immune antibodies; RhD-negative population, especially women should be highly aware of avoiding mis-transfusion of RhD-positive blood, and also avoiding multiple pregnancies resulting in newborn's hemolytic disease.
Base Sequence ; Blood Donors ; Exons ; Genotype ; Humans ; Isoantibodies ; Phenotype ; Rh-Hr Blood-Group System ; genetics ; immunology ; Rho(D) Immune Globulin

The Rh blood group system is one of the most complex of the known human blood group polymorphisms, including above 45 blood group antigens. Considerable progress has been made in our understanding of the molecular basis of Rh in the past 10 years. The bases of Rh inheritance, RH gene and its evolution, the structure and function of Rh complex, as well as nonerythroid Rh homolog, have been determined. Further improvements have been made in the technology of Rh genotyping. The review provides an update on the advance of Rh blood group to give information in the practice of forensic science.
Blood Group Antigens/genetics* ; Blood Proteins/immunology* ; DNA Fingerprinting ; Forensic Medicine/methods* ; Genotype ; Humans ; Molecular Biology ; Mutation ; Polymorphism, Genetic ; Rh-Hr Blood-Group System/genetics*

To investigate the RHD gene profiles of RhD-negative individuals in population of Fujian Province, it was to design fourteen pairs of specific primers to amplify RHD exon 1, 3 approximately 7, 9, 10, hybrid Rh box, RHD 1227A allele, RHC allele, RHc allele, RHE allele and RHe allele. Rh genotypes were detected by PCR-SSP in 104 RhD-negative donors, some samples with or without RHD genes were analysed by the absorption-elution test, and two RhD-negative samples with eight RHD exons detected were analysed by DNA sequencing. The results showed that 61.54% RhD-negative individuals lacked all the eight RHD exons detected (RHD-/RHD-), 25.97% carried the RHD 1227A allele (62.96% of which were the heterozygote of RHD+/RHD-, and 37.04% were the homozygote of RHD+/RHD+), 8.65% carried the RHD-CE (2 approximately 9)-D allele (RHD+/RHD-), and 1.92% carried the RHD 710delC allele (RHD+/RHD-). Though the most cases of RHD gene deletion were found in dce haplotype, six cases of RHD gene deletion were found in dCe (their RH genotypes were dce/dCe) and two in dcE haplotype (their RH genotypes were dce/dcE). And it was not accurate to predict the Rh phenotype by detecting a single RHD exon, however, and more accurate when eight RHD exons and RHD 1227A allele were detected (chi2=24.43, p<0.005). It is concluded that RHD genes in population of Fujian Province are polymorphic and the RHD genotyping is not reliable enough to replace the RhD serotyping in China.
Asian Continental Ancestry Group ; genetics ; Blood Donors ; China ; Erythrocytes ; immunology ; Exons ; genetics ; Gene Deletion ; Genotype ; Humans ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

This study was purposed to investigate the molecular basis for RhD negative phenotype in Yiwu population in Zhejiang Province of China. The RhD negative samples were screened by saline agglutination test in blood donors. Some real RhD negative and RhDel phenotypes were identified using anti-human globulin test and absorbtion elution test. Ten exons of RHD gene in these samples were amplified by PCR-SSP, and positive exons were DNA sequenced. The results indicated that 30 real RhD negative and 8 RhDel phenotypes were identified in 38 initial RhD negative samples. Ten exons were complete negative in 28 real RhD negative samples and only exon 1, 2 and 10 were positive in 2 real RhD negative samples amplified by PCR. All 10 exons in 8 RbDel samples were positive and a DNA variant (1227G > A) was found in 8 RhDel samples. It is concluded that all exons are absence in most real RhD negative phenotypes, and the partial exons absence is also found in some real negative phenotypes among Yiwu population in Zhejiang province of China. The G to A mutation at position 1227 is found in all RhDel phenotypes.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Exons ; Genotype ; Humans ; Molecular Sequence Data ; Phenotype ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics ; immunology

Extremely weak D variants called DEL are serologically detectable only by adsorption-elution techniques. A nucleotide change of exon 9 in RHD gene, RHD (K409K, 1227G>A) allelic variant is present in almost all the DEL individuals of East Asians. No DEL phenotype has yet been shown to induce a primary alloanti-D immunization in East Asia. A 68-yr-old D-negative Korean man was negative for anti-D at admission, and he developed alloanti-D after transfusion of red blood cells (RBC) from 4 apparently D-negative donors. Four donors who typed D-negative by routine serologic test were analyzed by real-time PCR for RHD gene and RHD (K409K). One donor was found to have RHD (K409K). This is the first case in which DEL RBCs with RHD (K409K) induced a primary alloanti-D immunization in Asian population. Because the DEL phenotype can induce an anti-D immunization in D-negative recipients, further discussion is needed whether RhD negative donors should be screened by molecular method and what an efficient genotyping method is for detecting the RHD gene carriers in Korea.
Aged ; Blood Donors ; Blood Grouping and Crossmatching ; Blood Transfusion/*adverse effects ; Exons ; Humans ; Isoantibodies/*metabolism ; Male ; Phenotype ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System/genetics/*immunology

Flow cytometry was previously applied for analysis of Rh(D) antigen density, therefore it was suggested that the flow cytometry may be used for routine detection of weak D positive phenotypes. This study was purposed to evaluate its practicability. Six weak D positive and 7 DEL individuals were detected by using saline, IAT and absorption/elution test from 2010 to 2011 years. By RHD genotyping, zygosity analysis and sequencing, 3 cases of weak D type 15, 3 cases of partial D type DVI-III and 7 cases of DEL carrying RHD1227A alleles were identified. Taking 2 normal Rh(D)-positive and 2 D-negative samples as controls, all the samples were tested by using flow cytometry, and the median fluorescence intensities were observed as well. The results indicated that all weak D type 15 and partial D type DVI samples were detected to be positive by flow cytometry, as compared with 2 Rh(D)-negative samples (P < 0.05). Seven 7 DEL samples were tested to be negative (P > 0.05), although one of 7 DEL was tested as "±" in IAT and strong positive in absorption/elution. The RHD zygosity analysis showed this DEL individual as RHD(+)/RHD(+) homozygote. It is concluded that the sensitivity of detecting D antigen by flow cytometry is similar to that of IAT, but lower than absorption/elution test. As for detecting weak D or partial D antigens, IAT is easier than flow cytometry; as for identifying DEL, the flow cytometry is not sensitive enough.
Adult ; Alleles ; Blood Donors ; Blood Grouping and Crossmatching ; Flow Cytometry ; methods ; Genotype ; Humans ; Phenotype ; Rh-Hr Blood-Group System ; blood ; genetics ; immunology

OBJECTIVETo report a rare case of hemolytic disease of the newborn (HDN) with kernicterus caused by anti-Di(a) diagnosed using high-throughput genotyping multiplex ligation-dependent probe amplification (MLPA).

METHODSConventional serological methods were used to detect the antibodies related with HDN. The genotypes of more than 40 red blood cell antigens for the newborn and her parents were obtained using the high-throughput MLPA assay. The antibody titers were tested using a standard serological method.

RESULTSThe unknown antibody against the low-frequency antigens was predicted based on the primary serological tests. The genotyping results for more than 40 red blood cell antigens of the newborn and her parents showed incompatible antigens of MNS and Diego blood group system, indicating the existence of anti-N or anti-Di(a). Further serological tests confirmed anti-Di(a) existence in the plasma of the newborn and her mother. The titer of anti-Di(a) in the mother's plasma was 1:32.

CONCLUSIONSevere HDN including kernicterus can result from anti-Di(a). High-throughput genotyping MLPA assay can help type some rare antigens in complicated cases. The reagent red cell panels including Di(a)-positive cells are necessary in routine antibody screening test in Chinese population.

Blood Group Incompatibility ; genetics ; Erythroblastosis, Fetal ; diagnosis ; immunology ; Exchange Transfusion, Whole Blood ; Female ; Genotype ; Humans ; Infant, Newborn ; Nucleic Acid Amplification Techniques ; methods ; Rh-Hr Blood-Group System ; genetics ; immunology ; Rho(D) Immune Globulin ; genetics ; immunology

This study was purposed to investigate the molecular basis of Rh DEL phenotype. Rh DEL phenotypes were identified by a serologic adsorption-elution method, the nucleotide sequences of ten RHD exons and exon-intron boundary regions were evaluated by a RHD gene-specific PCR-SSP (PCR-SSP, polymerase chain reaction-sequence specific primer) and sequencing. The results showed that out of 122 random Rh negative donors 35 Rh DEL phenotypes were identified through serologic method, including 6 RhCCdee (17.14%), 28 RhCcdee (80.00%), and 1RhCcdEe (2.86%). Sequence analysis indicated that all DEL phenotypes harbored a RHD 1227 G > A mutation in exon 9. D zygosity test revealed that 29 DEL phenotypes (28 RhCcdee and 1 RhCcdEe) had one RHD gene deleted, and 6 DEL phenotypes (6 RhCCdee) had homogenous RHD gene. It is concluded that RHD 1227A is an important genetic marker for Rh DEL phenotype in Zhejiang Han population.
Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Blood Donors ; China ; ethnology ; Erythrocytes ; immunology ; Exons ; genetics ; Humans ; Molecular Sequence Data ; Phenotype ; Point Mutation ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics ; immunology ; Sequence Analysis, DNA