The significance of detecting the recipient's Rh D blood type before transfusion and the feasibility of in-time transfusion to Rh D-negative person with the same ABO type blood or autologous blood were explored. For 2,168 inpatients who applied for transfusion for the first time in the latest five years, Rh D-antigen was detected a day before transfusion by test tube method with IgM anti-D serum provided by BASO company. The results showed that seven Rh D-negative cases were found among 2,168 cases and the Rh D-negative rate was 0.32%. Of these seven cases, two were received autologous blood, two were received Rh D-negative blood of the same type and the other three were not treated with transfusion. It is concluded that hemolytic reaction of transfusion was prevented by detecting Rh D blood type before transfusion, autologous transfusion may be taken if the patient's health-condition permits.
ABO Blood-Group System ; Adult ; Aged ; Blood Transfusion ; Female ; Humans ; Male ; Middle Aged ; Rh-Hr Blood-Group System ; analysis

BACKGROUND: Despite the advances in total laboratory automation, a considerable amount of work in blood banks is still done using outdated manual methods. Some automated pre-transfusion testing instruments have recently been developed. Of these, we evaluated and compared the AutoVue Innova (Ortho, USA) and the Techno TwinStation (DiaMed AG, Switzerland). METHODS: Forward and reverse ABO/Rh typing and unexpected antibody screening and identification tests were performed on 4,628 samples using the manual method and the two automated instruments. Two different anticoagulants (EDTA and citrate) were compared in ABO/Rh typing and unexpected antibody screening tests. Titrating studies were conducted on the following 7 dilutions using 5 samples of irregular antibodies with anti-E, anti-E & -c, anti-D, and anti-Le(a) with anti-Fy(a): 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, and 1:128. The test throughput per hour, the time required to perform 1 and 100 tests, and a simulation test for total events occurring in 1 day were also measured. RESULTS: No erroneous results were reported between the two instruments and the manual method. Discrepancies observed in 10 cases (0.4%) of ABO/Rh typing were of higher intensity with AutoVue Innova than with the manual method. AutoVue Innova exhibited the highest sensitivity in the titrating study and throughput performance compared with the manual method and the Techno TwinStation. Especially in the throughput and time required to complete 100 antibody screening tests, AutoVue Innova had a 3.3- and 3.5-fold higher performance, respectively, than Techno TwinStation. CONCLUSIONS: Because both of the two fully automated instruments (AutoVue Innova and Techno TwinStation) had high levels of accuracy and performance, it is expected that use of fully automated instruments will reduce human labor, turnaround time, and operator error in the blood bank.
ABO Blood-Group System/blood ; Antibodies/blood ; Automation ; Blood Grouping and Crossmatching/*instrumentation ; Blood Transfusion ; Cost-Benefit Analysis ; False Positive Reactions ; Humans ; Rh-Hr Blood-Group System/blood

There are 3' non-coding region, downstream Rhesus box, SMP1 gene et cetera. after RHD stop code. This study was intended to determine the sequence of 3' non-coding region. One pairs of primer was designed and then a polymerase chain reaction (PCR) was established for specific amplification of whole length of 3' non-coding region of RHD in 10 Rh-positive and 10 D(el) samples. The PCR products were purified and directly sequenced. The results showed that all Rh-positive and D(el) samples were identical, which revealed that there were 103 bp between 3'-end of RHD coding region and 5'-end of downstream Rhesus box. The D(el) samples showed the same result with the normal Rh-positive sample. It suggests that lower expression of D antigen in D(el) red cells does not associate with 3' non-coding region of D(el) gene.
3' Untranslated Regions ; genetics ; Base Sequence ; Humans ; Molecular Sequence Data ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

To study the method for Rhesus box test and its significance, the sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed according to RhD gene sequence; the upstream, downstream and hybrid Rhesus boxes were determined by PCP-SSP and mismatched PCR. The results showed that this method was confirmed by DNA Standard test. It was shown that in unrelative RhD positive individuals RHD(+)/RHD(-), RHD(+)/RHD(+) genotype accounted for 9.00%, 91.00% respectively, and in RhD negative individuals RHD(+)/RHD(-), RHD(+)/RHD(+), RHD(-)/RHD(-) genotype were 26.14%, 3.92%, 69.94% respectively. It is concluded that the method of Rhesus box test was confirmed to be reliable and can be used for the identification of RhD haplotype gene structure, as well as for study on inheritance, clinical transfusion and neonatal hemolytic diseases.
Base Sequence ; Haplotypes ; Heterozygote ; Homozygote ; Humans ; Polymerase Chain Reaction ; methods ; Rh-Hr Blood-Group System ; analysis ; genetics

The study was purposed to analyze DNA and allele structure of the partial D phenotypes D(Va) and D(VI) of the Rhesus blood group in Chinese. Through polymerase chain reaction (PCR) and direct genomic DNA sequencing, the RHD gene was detected in three weak D individuals identified serologically. The results showed that among the three weak D individuals, one was identified as partial D phenotype D(Va) (Hus) type and genotyped DccEe; another two were testified as D(VI) III type and genotyped DCcee. Moreover, the breakpoints of the replaced region by RHCE in D(Va) (Hus) were 5' end of the exon 5 and 3' end of the intron 5, and there were 7 novel polymorphisms in intron 5: 23-25(GCA)2, 98G>A, 168-169insG, 205-206insT, 494-495insA, 1256-1257insC, 1347G>T. In conclusion the whole exon 5 and intron 5 are replaced by RHCE in D(Va) (Hus) detected in Chinese.
China ; Exons ; genetics ; Genotype ; Humans ; Introns ; genetics ; Phenotype ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics ; Sequence Analysis, DNA

In the present day, pretransfusion tests include ABO and RhD grouping, antibody screening, antibody identification, and cross matching. Although error rates for these tests have decreased compared to those in the past, clerical errors still occur. When exposed to RhD positive RBCs, a RhD negative person can produce anti-D that causes a severe hemolytic disease of the fetus and the newborn in addition to hemolytic transfusion reactions. Therefore, administration of RhD positive RBCs to a RhD negative person should be avoided. We experienced a RhD negative patient who had been misidentified as positive and transfused 35 units of RhD positive RBCs eight years ago, but did not have detectable anti-D in present. The red cells of the patient showed no agglutination with the anti-D reagent and a negative result in the standard weak D test. The multiplex PCR with sequence-specific priming revealed that the patient was RhD negative.
*Blood Group Incompatibility ; Blood Transfusion ; Erythrocytes/*immunology ; Humans ; Isoantibodies/*analysis/immunology ; Male ; Middle Aged ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System/*analysis/immunology

Here we report a severe case of hemolytic anemia of the newborn with kernicterus caused by anti-Di(a) antibody. A full term male infant was transferred due to hyperbilirubinemia on the third day of life. Despite single phototherapy, the baby's total bilirubin had elevated to 30.1 mg/dL. After exchange transfusion, total bilirubin decreased to 11.45 mg/dL. The direct antiglobulin test on the infant's red cells was positive. The maternal and infant's sera showed a negative reaction in routine antibody detection tests, but were positive in Di(a) panel cells. The frequency of the Di(a) antigen among the Korean population is estimated to be 6.4-14.5%. Anti-Di(a) antibody could cause a hemolytic reaction against transfusion or hemolytic disease of the newborn. We suggest the need for reagent red blood cell panels to include Di(a) antigen positive cells in antibody identification test for Korean.
Alleles ; Bilirubin/blood ; Erythroblastosis, Fetal/*diagnosis/*immunology ; Humans ; Infant, Newborn ; Isoantibodies/*analysis ; Male ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System/*analysis/blood

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

BACKGROUND: This study compared the estimated costs and times required for ABO/Rh(D) typing and unexpected antibody screening using an automated system and manual methods. METHODS: The total cost included direct and labor costs. Labor costs were calculated on the basis of the average operator salaries and unit values (minutes), which was the hands-on time required to test one sample. To estimate unit values, workflows were recorded on video, and the time required for each process was analyzed separately. RESULTS: The unit values of ABO/Rh(D) typing using the manual method were 5.65 and 8.1 min during regular and unsocial working hours, respectively. The unit value was less than 3.5 min when several samples were tested simultaneously. The unit value for unexpected antibody screening was 2.6 min. The unit values using the automated method for ABO/Rh(D) typing, unexpected antibody screening, and both simultaneously were all 1.5 min. The total cost of ABO/Rh(D) typing of only one sample using the automated analyzer was lower than that of testing only one sample using the manual technique but higher than that of testing several samples simultaneously. The total cost of unexpected antibody screening using an automated analyzer was less than that using the manual method. CONCLUSIONS: ABO/Rh(D) typing using an automated analyzer incurs a lower unit value and cost than that using the manual technique when only one sample is tested at a time. Unexpected antibody screening using an automated analyzer always incurs a lower unit value and cost than that using the manual technique.
ABO Blood-Group System/blood ; Antibodies/analysis ; Automation ; Blood Banks/*economics/*standards ; Blood Grouping and Crossmatching/*economics/instrumentation ; Costs and Cost Analysis ; Humans ; Rh-Hr Blood-Group System/blood ; *Workflow ; Workload

OBJECTIVETo evaluate the status of eight RHD specific exons in 131 Han Chinese blood donors who were classified as RhD-negative by serological methods and explore the genomic structure of RHD gene among the Han Chinese. The Rh blood group system has the highest prevalence of polymorphisms among human blood group systems and is clinically significant in transfusion medicine. The Rh antigens are expressed on polypeptides encoded by two highly homologous genes, RHD and RHCE. Recent molecular studies have shown that the RhD-negative trait could be generated by multiple genetic mechanisms and is ethnic group-dependent.

METHODSThe polymerase chain reaction using-sequence specific primers (PCR-SSP) was used to amplify exons 2, 3, 4, 5, 6, 7, 9 and 10 of RHD gene and exons 1, 2 and 5 of RHCE gene, as well as intron 4 in each of them.

RESULTSThe 131 cases of RhD-negative phenotypes consisted of 60 ccee, 58 Ccee, 5 ccEe, 5 CcEe and 3 CCee. Among them, 83 with the Rh ccee or ccEe phenotypes (63.4%) lacked the eight RHD exons indicated above, while 26 cases with the Rh Ccee, CCee, CcEe phenotypes (19.9%) had all the RHD exons examined. Twenty-two individuals with the Ccee, CCee, CcEe phenotypes (16.8%) carried at least one RHD exon. The phenotypes of the RhD negative individuals carrying the RHD gene were Rh CC or Cc, but not cc.

CONCLUSIONSThree classes of RhD-negative polymorphisms among a population of Han Chinese were observed. Antigen association analysis suggested the existence of a novel class of RhD-negative associated haplotype in Han Chinese. This haplotype consisted of a normal RHCE allele and a nonfunctional RHD gene. It may be beneficial to redefine the RhD-negative blood group among Chinese populations upon clarification of the mechanisms of RHD gene expression and RhD antigen immunization.

Asian Continental Ancestry Group ; China ; Ethnic Groups ; genetics ; Humans ; Phenotype ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; analysis ; genetics