Discussion on the collection and supply mode of RhD negative blood suitable for prefecture-level city: a retrospective analysis / 中国输血杂志

【Objective】 To explore a RhD negative blood collection and supply mode suitable for Liaocheng area and improve the blood demand satisfaction rate of RhD negative patients. 【Methods】 Under different collection and supply modes (collection mode: in 2020, not advocate walk-in donation by RhD negative repeat blood donors, but in 2021 and 2022 were the opposite; supply mode: in 2020, type A, B, O and AB RBCs were frozen within 6 days of blood collection without inventory, but in 2021 and 2022, they were stored of 2-6 U and the remaining were frozen within 6 days of blood collection), RhD negative blood was divided into Type A, B, O and AB, appointment donation and walk-in donation, first donation and repeat donation, cold storage red blood cells(RBCs), frozen RBCs and frozen thawed deglycerolized RBCs, and the collection and supply data of each observation group from 2020 to 2022 were compared. Based on whether blood demand of RhD negative patients was met, the patients were divided into the group of going to other places for medical treatment, the group of RhD positive blood transfusion and the group of RhD negative blood transfusion to analyze the blood usage. 【Results】 From 2020 to 2022, the proportion of RhD negative blood donated by repeat appointment donors decreased year by year (P<0.05); the proportion of RhD negative blood donated by repeat walk-in donors increased year by year (P<0.05); the proportion of frozen thawed deglycerolized RBCs to RhD negative RBCs increased year by year (P<0.05); the proportion of cold storage RBCs distributed as RhD negative and RhD positive decreased year by year respectively (P<0.05); the proportion of the increase in frozen RBCs inventory to the current year's frozen RBCs inventory decreased year by year (P<0.05). The satisfaction rate of blood demand of RhD negative patients increased year by year (P<0.05), with the number of patients with RhD positive blood transfusion and going to other places for medical treatment decreased year by year respectively(P<0.05). 【Conclusion】 In Liaocheng area, it is appropriate to encourage RhD negative blood donors to donate blood randomly, and A, B, O and AB type cold storage RBCs kept in 2-6 U inventory respectively. When exceeding the inventory, frozen RBCs are prepared within 6 days, which can improve the blood demand satisfaction rate of RhD negative patients and avoid sending excessive RhD negative RBCs to clinical use as RhD positive RBCs.

Molecular basis of RhD-negative phenotype in North Indian blood donor population

Background & objectives: RHD gene typing is highly complex due to homology with RHCE genes. Molecular polymorphism of the RHCE and RHD genes have been characterized among various populations, but no studies have been undertaken among Indians. This study was undertaken to assess the genetic basis of RHD-negative phenotype in Indian blood donor population. Methods: Sample from a total of 200 phenotypically RhD-negative blood donors were analyzed for presence of RHD gene using polymerase chain reaction (PCR). RHD genotyping was done using three primer sets designed for exons 4 and 10 and one set for identification of pseudo (RHD?) gene between introns (int) 3 and 4. Amplified PCR products were analyzed by gel-electrophoresis (XY Loper, Uvitech, Cambridge) and confirmed by nucleotide sequencing (ABI 3730 xl 96 capillary system). Results: No PCR product was found in 195/200 (97.5%) of study samples indicating homozygous gene deletion. Of the 5/200 (2.5%) showing RHD gene polymorphisms, 4/200 (2%) were positive for presence of exon 10 only (RHD-CE-D hybrid). RHD? gene was not detected in any of the samples tested. One sample showed presence of all three tested regions and was negative for RHD? gene. Interpretation & conclusions: RHD gene deletion was found to be the most common cause of an RHD-negative phenotype while RHD? gene was, reported to be present in up to 39 per cent of various ethnic populations, but was not detected. RHD-CE-D hybrid gene (found in 2.5% individuals) is important for predicting the requirement of Rh prophylaxis during the antenatal period.

Phenotype and RHD gene polymorphism of 64 Rh negative individuals / 中国输血杂志

【Objective】 To collect blood samples of 64 RhD negative patients in our hospital for RHD genotyping and phenotype analysis (RhC/c/E/e), and analyze the distribution characteristics of different RHD genotypes. 【Methods】 The RHD gene of RhD negative patients was genotyped by fluorescence quantitative polymerase chain reaction (PCR) method. The Rh phenotype was identified by IgM anti-e, anti-c, anti-C and anti-E, respectively. 【Results】 Forty-two cases of RHD deletion were detected, dominated by ccee phenotype (88.1%); 9 of RHD1227A cases, dominated by Ccee phenotype(77.8%); 8 of RHD-CE(3-9)-D2 cases, dominated by Ccee phenotype (75%); 1 of RHD-CE(3-10)-D2 case with Ccee phenotype, 1 of RHD*711delC case; 1 of RHAG site invalid type were detected. The typing results could not be determined in 2 cases by PCR method. 【Conclusion】 RhD negative patients showed diversity in RHD genotype, dominated by RHD deletion, followed by RHD1227A, RHD-CE(2-9)-D2, RHD-CE(3-10)-D2, RHD*711delC and RHAG site deletions.

Molecular basis of Del phenotype in RhD negative population of Zhongshan / 中国输血杂志

【Objective】 To conduct serological and molecular study of Del type in RhD-negative donor population in Zhongshan area, so as to improve the diagnosis of Del type. 【Methods】 A total of 102 initially RhD-negative samples, collected from December 2017 to February 2019, were classified by RHCE and PCR-SSP genotyping. And 95 cases of truly negative RhD were confirmed by IAT, 28 cases of Del type were identified by absorption and elution test. The phenotype and genotyping characteristics of Del type in Zhongshan area were summarized based on domestic data of relative literature. 【Results】 Among 102 initially RhD-negative samples by serological test, 95 were truely RhD-negative, 28 were DELRHD 1227A without any other Del allele. Among them, RHCE antigen type were Ccee in 20(71.4%) cases, CCee in 8(28.6%), with no difference in comparison with other regions in China. The frequency of Del in RhD-negative blood donors was 29.5% (28/95), with difference between Shanghai, Taiwan, and Fuzhou, but no difference between Nanchang, Zhejiang, and Wuhan. 【Conclusion】 The study showed that the Del phenotype was closely related to Ce haplotype, and has no difference with other regions in China. The frequency of Del type in RhD negative donors was 29.5%, with regional differences. RHD1227A was the main allele of Del.

Study on the gene frequency and polymorphism of 11 RBC blood group systems in RhD negtive population in Hunan, China / 中国输血杂志

【Objective】 To investigate the gene frequency and polymorphism of RBC blood group systems in RhD negtive population in Hunan, so as to lay a foundation for clinical blood transfusion and construction of multiple rare blood group database. 【Methods】 Blood samples were taken from 300 RhD negative blood donors, confirmed by serological method, from June 2019 to June 2020,. RHD genotyping was performed by SSP-PCR. For blood donors with typing results as RhD negative plus RHD gene deletion, antigens genotyping of MNS, Duffy, Kell, Domrock, Diego, Kidd, Sciawnna, Colton, Lutheran and Yt RBC blood group systems were performed by SSP-PCR and analyzed by the chi square test of SPSS 20 statistical software. 【Results】 RHD gene deletions accounted for 58.67% (176 / 300) of serological D negative blood donors. The gene frequencies were as follows: MNS: GYPB*S=0.045 5(8/176), GYPB*s=0.954 5(168/176), GYP*Dane=0.039 8(7/176); Duffy: FY*A =0.965 6(170/176), FY*B=0.034 1(6/176); Dombrock: DO*A=0.082 4(14.5/176), DO*B=0.917 6(161.5/176); Diego: DI*A=0.025 6(4.5/176), DI*B =0.974 4(171.5/176); Kidd: JK*A=0.485 8(85.5/176), JK*B=0.514 2(90.5/176); Kell: KP*A=0.005 7(1/176), KP*B=0.994 3(175/176); Lutheran: LU*A=0.005 7(1/176), LU*B=0.994 3(175/176); Yt: YT*A=0.002 8(0.5/176), YT*B=0.997 2(175.5/176). The genotypes of Kell(K+ /k+ ), Scianna and Colton blood groups were KEL*02 /KEL*02, SC*01 /SC*01 and CO*A /CO*B, respectively. The expected frequencies of the combination of type O, RhD negative and other blood group systems were between 1/100 000 to 1/10 000. 【Conclusion】 Among RhD negative blood donors in Hunan, the gene profiles of MNS, Duffy, Domrock, Diego, Kidd, Kell and Lutheran blood group system were polymorphic, and Kell (K+ /k+ ), Colton and Scianna were homozygous. The data of other RBC blood group systems from RhD negative blood donors is of great significance to establish local database of rare blood groups.

Genetic background of RhD negative blood donors / 中国输血杂志

【Objective】 To analyze the genetic background of RhD-negative blood donors by detecting RHD and RHCE genes of those donors. 【Methods】 From March 2021 to May 2022, the blood samples of RhD-negative blood donors, who had been screened out by RhD primary screening and confirmatory experiments in the Yaan Blood Center, were firstly identified whether the RHD allele was completely deleted, then whether there were deletions in 10 exons of non-RHD allele complete deletion samples, finally, the remaining samples without RHD alleles and exon deletions were further analyzed by DNA sequencing. RHCE gene was detected by SSP-PCR method. 【Results】 Among the RHD gene test results of 104 RhD-negative samples, 65 cases were completely deleted (d/d), 33 were RHD partially deleted (one allele deletion), and 6 were without RHD gene deletion. The RHD alleles of 33 samples with partial deletion were detected by 10 exons, 13 had partial exon deletion, with genotype as RHD*D-CE(3-9)-D/d and phenotype as RhD negativity, and the remaining 20 samples had no exon deletion. The exon sequencing results of the non-deletion samples showed RHD*1227A/RHD*1227A in 6 samples, RHD*1227A/d in 19, RHD*3A/d in 1; both of the last two were considered Del by ISBT. The RHCE gene test results showed that all cc genotype blood donors were RhD true negative, while Del blood donors had no cc genotype. 【Conclusion】 Through the genetic background study of RhD negative blood donors, it is found that there is a high proportion of Del with weak expression of RhD antigen, whether this blood type affects clinical blood safety needs further researches.

Safety of RhD alloimmunization / 中国输血杂志

【Objective】 To explore the safety of RhD-positive red blood cells (RBCs) immunization schedules in RhD-negative volunteers, so as to facilitate the development of domestic anti-D immunoglobulin. 【Methods】 From January 2018 to April 2020, 23 RhD negative volunteers with informed consent were enrolled and divided into initial immunization group and booster immunization group. The initial immunization included first immunization, second immunization and third immunization. Four groups, i. e. 3 cases of 20 mL, 8 of 30 mL, 6 of 40 mL, and 6 of 50 mL, were involved in initial immunization. After the initial immunization response, booster immunizations were performed every 3 months. According to the anti-D titer before each immunization, the booster immunization doses were set to 0.5, 1 and 2 mL. Whole blood samples of 5mL/ person (time) were collected 24 h and 1 week after each infusion, and the blood routine, liver, kidney and blood coagulation function and anti-D titer were detected. The differences of detection (index) values at 24 h and 1 week after the first immunization and booster immunization in each (dose) group were compared. 【Results】 No statistically significant differences were observed in hemolysis index values (all within the range of medical reference values) 24 h or 1 week after initial immunization among RhD positive RBCs of 20, 30, 40 and 50mL(P>0.05). The differences between the hemolysis index values and the basic values before the immune response (all within the range of medical reference values) after 0.5 or 1 mL booster immunizations were also not statistically different (P>0.05). However, the differences (μmol/L)between total bilirubin levels and the basic values before the immune response (1.55±1.87, 6.29±2.66) were significantly different after 2 mL booster immunization (P<0.05). 【Conclusion】 No risks affecting the safety of RhD negative volunteers was found in the immunization schedule proposed in this study.

Genetic polymorphism and antibody screening of RhD negative Chinese Han population in Jiayuguan / 中国输血杂志

【Objective】 To investigate and analyze the polymorphism of RHD gene in RhD-negative population in Jiayuguan using molecular biological technique, so as to accurately identify RhD-negative individuals, and formulate individualized transfusion strategies. 【Methods】 The RhD negative voluntary blood donors and patients (mainly pregnant women) were recruited. After informed consent, history of blood transfusion and pregnancy of them were investigated, and samples were collected for negative D confirmation, gene sequencing as well as antibody screening and identification. 【Results】 Among the 96 samples, 73 cases were RHD gene deletion, 18 RHD*01EL.01(17 RHD1227A homozygous type and 1 RHD1227A heterozygous type), 2 weak RHD*15 type (845G/A), 1 partial D type, i. e. RHD-CE(7) -D heterozygous allele, and 2 RHD*01N.16 variant. Antibody was detected out in 4 cases, among which 2 were positive for anti-D, 1 anti-D plus anti-E, and 1 anti-Dia. 【Conclusion】 The proportion of DEL gene in RhD negative Chinese Han population in Jiayuguan is slightly lower than that in general Chinese Han population. No anti-D or RHD-HDN was observed in DEL type women due to multiple pregnancy or delivery of D positive newborns.

Severe anti-D haemolytic disease of fetal and newborn in rhesus D negative primigravida

@#Introduction: Anti-D alloimmunisation may occur from the blood transfusion or fetomaternal haemorrhage which can lead to haemolytic disease of fetal and newborn (HDFN). The morbidity and mortality of HDFN related to anti-D is significantly reduced after introduction of anti-D prophylaxis and furthermore, anti-D HDFN in RhD negative primigravida is uncommonly seen. Case Report: A case of unusual severe HDFN due to anti-D alloimmunisation in undiagnosed RhD negative primigravida Malay woman is reported here. This case illustrates the possibility of an anamnestic response from previous unknown sensitisation event or the development of anti-D in mid trimester. The newborn expired due to hydrops fetalis and severe anaemia. Antenatally, the mother was identified as RhD positive and thus there was no antenatal antibody screening, antepartum anti-D prophylaxis or close fetal monitoring for HDFN. Discussion: The thorough antenatal ABO and RhD blood grouping with antibody screening is mandatory as part of prevention and early detection of HDFN especially due to anti-D alloimmunisation. Improper management of RhD negative women might lead to severe HDFN including in primigravida.

Analysis on effect of blood anti-D antibody detection in infusing RhD positive homotype RBC for patients with RhD negative / 国际检验医学杂志

Objective To investigate the anti-D antibody level after infusing homotype RhD positive RBC in the patient with RhD negative.Methods The clinical data in 20 cases of RhD negative infusing homotype RhD positive RBC in our hospital from January 2010 to January 2016 were collected.The anti-D antibody levels before blood infusion and on 10,20,30,90 d after blood infusion and the titers in the patients with RhD positive were analyzed.Results Among 20 cases of RhD negative infusing RhD positive homotype RBC blood,6 cases were RhD positive within 90 d after blood infusion,in which the RhD positive rates were 25%(3/12) in male and 37.5%(3/8) in female.The anti-D antibody titers in 3 cases of RhD positive were 35,278 and 508 respectively.Conclusion Infusing RhD positive RBC blood in the patients with RhD negative can stimulate the immune mechanism,generates the anti-D antibody at RBC surface.Moreover the RBC phenotype in partial patients with RhD negative may change.

Identification of Anti-G in Pregnant Women with RhD Negative Blood: The First Case in Korea / 대한수혈학회지

Anti-G positivity can be misinterpreted as the presence of anti-D or -C antigen in an antibody identification test, as this antibody is known to show agglutination to D or C antigen-positive red cells. Correct identification of anti-G is important in pregnant women, as prenatal care or the need for RhIG administration can vary between anti-D and -C versus anti-G cases. We recently encountered a D-negative case with suspected anti-D and -C, which was ruled out by adsorption and elution tests, and ultimately confirmed the presence of anti-G. The pregnant woman was a 33-year-old patient with cde Rh phenotype with a previous history of spontaneous abortion, followed by administration of RhIG. The spouse's Rh phenotype was CDe. Initial antibody identification test showed 2+ positivity to C (homozygotes and heterozygotes) and trace to 1+ positivity to D. Upon additional adsorption and elution with R0r (cDe/cde) and r'r (Cde/ cde) red cells, we identified the antibody present in the patient's serum as anti-G. The patient is currently under close follow-up monitoring for anti-G titer using antibody titer testing with both CDe and CcDEe red cells. Periodic fetal cerebral Doppler examination is being carried out without evidence of fetal distress.

Rh Blood Grouping of the Individuals Born of Rh Negative Mother and Rh Positive Biological Father.

Unlike ABO antigens, Rh antigens are present only on Red Blood Cells (RBC), RhO(D) antigen is clinically the most important in the Rh system because it is highly antigenic. In India, as reported, 95% population is Rh positive whereas 05% is Rh Negative(approximately). Rh antibodies are the major cause3 of haemolytic disease of newborn (HDN) and lead to destruction of transfused Rh Positive red cells. A total number of 799 pregnant mothers having RhD Negative Blood Group (and also their husband possessing RhD Positive Blood Group) were surveyed. Out of the 799 babies born, 662 possessed RhD Positive and 137 possessed RhD Negative Blood Group. Besides this, twin babies born of RhD Negative mother and RhD Positive Biological Father were computed separately (Total No.of such mothers being 12). In case of twin delivery, the twins babies all possessed RhD Positive Blood Group. Calculation shows that about 17% of babies were RhD Negative and 83% were RhD Positive.

Liver transplantation between a RhD positive graft to a RhD negative recipient / 中华肝胆外科杂志

ObjectiveTo investigate the choice of graft,and transfusion and immunosuppressant regimen of a RhD negative recipient in liver transplantation.MethodsOne RhD negative patient with hepatocellular carcinoma who received a liver graft from a RhD positive donor was retrospectively studied,and related references were reviewed.During the operation,the patient received five units of RhD negative/O RBC,3000 ml positive/O plasma and 30 units cryoprecipitate.Tacrolimus and prednisone were used to prevent rejection,and prednisone was withdrawn 30d post transplant.Results The patient's liver function recovered smoothly,without any acute rejection or hemolytic reaction.Anti-D antibody was not detected.The patient suffered from cancer recurrence 9 months and died of brain metastasis 13 months after transplantation.ConclusionsA RhD negative recipient can receive a graft from a RhD positive donor in liver transplantation.The selection of RBC and platelet from RhD negative or positive donors should be based on the result of anti-D antibody test.Plasma and cryoprecipitate can be transfused regardless of Rh type.Enhanced immunosuppressant regimen was unnecessary for these patients.

Alelos RHD nulos en pacientes RhD negativo de una población hospitalaria de la ciudad de Rosario / RHD null alleles in RhD negative patients from a hospital of Rosario

El fenotipo RhD negativo en la población caucásica es causado por una deleción completa del gen RHD. Sin embargo, han sido reportadas regiones específicas de este gen en individuos RhD negativo de diferentes grupos étnicos. El objetivo de este trabajo fue investigar la presencia de alelos RHD nulos en pacientes RhD negativo que concurrieron al Hospital Provincial del Centenario. Se tipificaron 12672 individuos y se seleccionaron las muestras RhD negativo halladas. Se determinó el fenotipo Rh completo y posteriormente se investigó la presencia del gen RHD utilizando una estrategia de PCR multíplex. En las muestras que presentaron fragmentos RHD específicos se realizaron reacciones de PCR alelo específicas para determinar el origen de los exones. Se encontraron 653 (5.15%) muestras RhD negativo. Cincuenta y cinco (8.42%) presentaban al menos el antígeno RhC o RhE. Los estudios moleculares permitieron detectar 7 alelos RHD Psi, 5 alelos híbridos RHD-CE(3-7)-D, 2 alelos híbridos RHD­CE(3-9)-D y 1 alelo nuevo RHD (46 T>C). La frecuencia de individuos RhD negativo en la población estudiada fue significativamente menor a la reportada en caucásícos. Los resultados moleculares obtenidos indican que 2.30% (15/653) de los individuos que no expresan el antígeno D son portadores de alelos RHD nulos. Los alelos RHD-CE(3-7)-D, RHD-CE(3-9)-D y RHD (46 T>C) están presentes únicamente en individuos RhD negativo que expresan los antígenos RhC y/o RhE con una frecuencia del 14.50% (8/55). Por otro lado, el alelo RHD Psi está asociado exclusivamente al fenotipo dccee, siendo el 1.17% (7/598) de estos individuos portadores del pseudogen RHD Psi. Estos hallazgos señalan la importancia del estudio del polimorfismo molecular del locus RH para el desarrollo de estrategias de tipificación de ADN confiables, que permitan realizar la genotipificación RHD prenatal y optimizar la selección de unidades a transfundir en los Bancos de Sangre.

The RhD negative phenotype in Caucasians is mainly caused by a complete deletion of the RHD gene. However, specific regions of the RHD gene in RhD negative individuals have been reported in different ethnic groups. The purpose of this study was to analyse the presence of silent RHD alleles in RhD negative patients concurring to the Hospital Provincial del Cen­tenario. Blood samples from 12672 individuals were studied and the RhD negative phenotypes were selected. Initially, the complete Rh phenotype was determined and DNA samples were screened using a multiplex PCR strategy to detect the presence of an RHD allele. Samples carrying RHD specific fragments were further studied by RHD exon scanning with allele specific PCR. 653 samples out of the 12672 (5.15%) were found RhD negative. Within this group, 8.42 % were either RhC positive or RhE positive. Molecular studies detected 7 RHD Psi alleles, 5 RHD-CE(3-7)-D hybrid alleles, 2 RHD-CE(3-9)-D hybrid alleles and 1 RHD (46 T>C) novel allele.The frequency of RhD negative individuals observed in the population studied was lower than that reported for Caucasians. Molecular analysis showed that 2.30% (15/653) of the individuals with no expression of the D antigen carry RHD null alleles. RHD­CE(3-7)-D, RHD-CE(3-9)-D and RHD (46 T>C) alleles are present only in individuals expressing either RhC or RhE with a frequency of 14.55% (8/55). The RHD Psy is associated with the dccee phenotype and 1.17% (7/ 598) of these individuals carries the RHD Psi pseudogen. These findings highlight the importance of studying the molecular polymorphism of the RH locus so as to develop reliable DNA typing strategies.

A RhD Negative Patient Failed to Produce Detectable Anti-D after Transfusion of 35 Units of RhD Positive Red Blood Cells / 대한진단검사의학회지

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.