Establishment of a genotyping method for the junior blood group and identification of a rare blood type with partial DVI.3 and Jr(a-) / 中华医学遗传学杂志

OBJECTIVE@#To develop a genotyping method for the Junior blood type and report on a rare blood type with Jr(a-).@*METHODS@#Healthy O-type RhD+ volunteer donors of the Shenzhen Blood Center from January to May 2021 (n = 1 568) and a pedigree with difficult cross-matching (n = 3) were selected as the study subjects. Serological methods were used for proband's blood type identification, unexpected antibody identification, and antibody titer determination. Polymerase chain reaction-sequence specific primer (PCR-SSP) method was used for typing the proband's RhD gene. ABCG2 gene coding region sequencing and a PCR-SSP genotyping method were established for determining the genotypes of the proband and his family members and screening of Jra antigen-negative rare blood type among the 1 568 blood donors.@*RESULTS@#The proband's ABO and RhD blood types were respectively determined as B and partial D (RHDDVI.3/RHD01N.01), Junior blood type Jra antigen was negative, and plasma had contained anti-D and anti-Jra. Sequencing of the ABCG2 gene revealed that the proband's genotype was ABGG201N.01/ABGG201N.01 [homozygous c.376C>T (p.Gln126X) variants], which is the most common Jr(a-) blood type allele in the Asian population. Screening of the voluntary blood donors has detected no Jr(a-) rare blood type. Statistical analysis of the heterozygotes suggested that the allelic frequency for ABCG2*01N.01 (c.376T) was 0.45%, and the frequency of Jr(a-) rare blood type with this molecular background was about 0.2‰.@*CONCLUSION@#A very rare case of partial DVI.3 type and Jr(a-) rare blood type has been identified. And a method for identifying the Junior blood type through sequencing the coding regions of the ABCG2 gene and PCR-SSP has been established.

Genotipificación del gen RHD en ADN fetal libre en plasma de embarazadas RhD negativo / Genotyping of the RHD gene in free fetal DNA in plasma of RhD negative pregnant women

Objetivo: Determinar el grupo RhD fetal a través del estudio del gen RHD en ADN fetal que se encuentra libre en plasma de embarazadas RhD negativo. Método: Se analizó la presencia de los genes RHD, SRY y BGLO en ADNfl obtenido de plasma de 51 embarazadas RhD negativo no sensibilizadas, utilizando una qPCR. Los resultados del estudio genético del gen RHD se compararon con el estudio del grupo sanguíneo RhD realizado por método serológico en muestras de sangre de cordón, y los resultados del estudio del gen SRY fueron cotejados con el sexo fetal determinado por ecografía. Se calcularon la sensibilidad, la especificidad, los valores predictivos y la capacidad discriminativa del método estandarizado. Resultados: El gen RHD estaba presente en el 72,5% de las muestras y el gen SRY en el 55,5%, coincidiendo en un 100% con los resultados del grupo RhD detectado en sangre de cordón y con el sexo fetal confirmado por ecografía, respectivamente. Conclusiones: Fue posible deducir el grupo sanguíneo RhD del feto mediante el estudio del ADN fetal que se encuentra libre en el plasma de embarazadas con un método molecular no invasivo desarrollado y validado para este fin. Este test no invasivo puede ser utilizado para tomar la decisión de administrar inmunoglobulina anti-D solo a embarazadas RhD negativo que portan un feto RhD positivo.

Objective: To determine the fetal RhD group through the study of the RHD gene in fetal DNA found free in plasma of RhD negative pregnant women. Method: The presence of the RHD, SRY and BGLO genes in fetal DNA obtained from plasma of 51 non-sensitized RhD negative pregnant women was analyzed using qPCR. The results of the genetic study of the RHD gene were compared with the RhD blood group study performed by serological method in cord blood samples, and the results of the SRY gene study were compared with the fetal sex determined by ultrasound. Sensitivity, specificity, predictive values and discriminative capacity of the standardized method were calculated. Results: The RHD gene was present in 72.5% of the samples and the SRY gene in 55.5%, coinciding 100% with the results of the RhD group detected in cord blood, and with the fetal sex confirmed by ultrasound, respectively. Conclusions: It was possible to deduce the RhD blood group of the fetus through the study of fetal DNA found free in the plasma of pregnant women with a non-invasive molecular method developed and validated for this purpose. This non-invasive test can be used to make the decision to administer anti-D immunoglobulin only to RhD-negative pregnant women carrying an RhD-positive fetus.

RHD Gene Analysis of A Blood Donor with Del Phenotype / 中国实验血液学杂志

OBJECTIVE@#To analyze the RHD genotype of a blood donor with Del phenotype in Yunnan.@*METHODS@#Rh serological phenotype was identified. RHD gene was detected by PCR-SSP typing, and its 10 exons were sequenced. Exon 9 was amplified for sequencing and analysis. RHD zygosity was detected.@*RESULTS@#The Rh phenotype of this specimen was CcDelee. Genomic DNA exhibited a 1 003 bp deletion spanning from intron 8, across exon 9 into intron 9. The deletion breakpoints occurred between two 7-bp short tandem repeat sequences. There was no variation in the sequences of the remaining exons. The Rh hybridization box test showed that there was one RHD negative allele.@*CONCLUSION@#This specimen is Del type caused by deletion of RHD exon 9.

Molecular Mechanism of a Rhesus D Variant Individual with RHD*845A/1227A / 中国实验血液学杂志

OBJECTIVE@#To explore the genetic mutation mechanism of a rare Rhesus D variant individual.@*METHODS@#Regular serological assay was used for determination of Rh type for the sample. Indirect anti-human globulin test (IAT) was used to confirm the RhD antigen and screen the antibodies. D-screen reagent was used to analyze the RhD epitopes of the sample. RHD genotype and RHD zygosity testing of the sample were detected by palymerase chain reaction with sequence-specific primers (PCR-SSP). The full length coding region of RHD gene was sequenced. RHD mRNA was detected using reverse transcription polymerase chain reaction (RT-PCR). The PCR products were cloned and sequenced.@*RESULTS@#The RhD blood group of the sample was determined as weak D, and the Rh phenotype was CcDEe. The antibody screening was negative. The sample tested with all monoclonal anti-Ds in D-screen showed the D epitope profiles as partial D types. The analysis of RHD gene sequence indicated that the individual with RHD c.845G/A and RHD c.1227G/A base heterozygosis. Three kinds of alternative splicing isoforms were obtained by TA cloning and sequencing.@*CONCLUSION@#The object has RHD c.845G/A and RHD c.1227G/A mutation. This heterozygous mutation is responsible for the low expression of RhD antigen on the red blood cells of the sample.

Investigation of Antigen and Gene Frequency of Kell(K) and Rh(D) Blood Groups in Xinjiang / 中国实验血液学杂志

OBJECTIVE@#To investigate the phenotypes and gene frequencies of Kell blood group system K antigen and Rh blood group system D antigen in Xinjiang, and summarize and understand the distribution of Kell(K) blood type and Rh(D) blood type in this area.@*METHODS@#A total of 12 840 patients who met the inclusion criteria during physical examination and treatment in our hospital and 18 medical institutions in our district from January 1, 2019 to December 31, 2019 were collected for identification of Kell blood group system K antigen and Rh blood group System D antigen, and the distribution of K and D blood groups in different regions, genders and nationalities were investigated and statistically analyzed.@*RESULTS@#The proportion of K positive in the samples was 1.39%, the highest was 1.91% in southern Xinjiang, and the lowest was 1.03% in northern Xinjiang(P<0.01). The proportion of Rh(D) negative samples was 2.75% and the gene frequency was 16.64%. The proportion of Rh(D) negative samples was 4.03% and the gene frequency was 20.10% in southern Xinjiang, followed by eastern Xinjiang and the lowest in northern Xinjiang (P<0.01). The frequency of K antigen in Uygur nationality was the highest, reaching 2.16%, Kirgiz 1.54%, and the distribution trend of D/d antigen was similar to that of K antigen. Among women, the K positive frequency of Kazak nationality was slightly higher than that of Mongolian nationality. The highest proportion of K positive in Uygur women was 2.38%, which was higher than that in Uygur men (1.86%). The frequency of d phenotype in Kazak women was 3.15%, which was higher than that in Kirgiz （2.89%） (P<0.01).@*CONCLUSION@#The distributions of Kell(K) and Rh(D) blood groups in northern and southern Xinjiang and eastern Xinjiang had its own unique characteristics and differences. There are significant differences in blood group distribution among different ethnic groups and gender groups. In the future, k antigen detection can be included to further improve the investigation on the distribution of Kell blood group system in this region.

Identification of Weak D Type 1 in Rh Blood Group System and Discussion of Transfusion Strategy / 中国实验血液学杂志

OBJECTIVE@#To investigate the molecular mechanism of one patient with abnormal serological phenotype in RhD and discuss the transfusion strategy.@*METHODS@#The RhD variant sample was screened from a patient with IgM type anti-D antibody and further determined by three different sources of anti-D antibodies. Ten exons and the adjacent introns of the RHD gene were amplified, purified and sequenced. RhCE phenotypes and RHCE genotypes were detected.@*RESULTS@#The patient with Rh variant showed abnormal results of serological tests. The RHD gene sequence analysis showed that the RHD*01W.01 with a variation (c.809T>G, p.Val270Gly) in exon 6 of the RHD gene was found in the patient. The RhCE phenotype was CcEe. The genotyping results of RHCE were consistent with the serological typing results.@*CONCLUSION@#The Rh variant of the patient is RHD*01W.01, these findings indicate that RhD variants should be analyzed by molecular assays for the sake of safe transfusion.

A pedigree analysis of a rare RhD 336-1G>A intron variant / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular mechanism of a case where RhD genotyping did not match serological results.@*METHODS@#The serological results of 8 members from two generations of this family were analyzed. And according to Mendelian law of inheritance, RhD genotyping, zygotic type determination and gene sequencing were performed for the family members.@*RESULTS@#The proband and one of her cousins have the same RhD alleles, both of them have a 336-1G>A intron variant RhD allele and a complete RhD deletion allele. The variant alleles are inherited from two of their parents with blood relationship, while the complete-deleted alleles come from the other. 336-1G>A means that the last base G of the second intron of the RhD gene is mutated to A, which leads to a negative RhD serology and a positive genotype in the proband.@*CONCLUSION@#There was a rare 336-1G> A intron variant gene (RhD * 01N.25) in this family, which was a recessive gene relative to the RhD gene and resulted in RhD phenotype negative.

Molecular genetic analysis of two individuals with weak D variant of the Rh blood type / 中华医学遗传学杂志

OBJECTIVE@#To explore the molecular basis of two individuals with weak D variant of the Rh blood type.@*METHODS@#Routine serological testing was carried out to detect the D, C, c, E and e antigens of the Rh blood group. The D antigen was further detected with an indirect antiglobulin test. The presence of Rhesus box was detected by PCR to determine the homozygosity of the RHD gene.@*RESULTS@#Both samples were determined as weak D phenotype by the indirect antiglobulin test. DNA sequencing revealed that case 1 harbored a heterozygous 208C>T variant in exon 2 and a heterozygous 1227G>A variant in exon 9; while case 2 harbored homozygous 779A>G variants of exon 5 of the RHD gene. Case 1 was determined as RHD+/RHD+, while case 2 was determined as RHD+/RHD-. The two samples were respectively named as weak D type 122 and weak D type 149 based on the rules of Rhesus Base Nomenclature.@*CONCLUSION@#D negative blood donors should subject to indirect antiglobulin testing and molecular analysis for safer transfusion.

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.

Weak D antigen - Revisited.

D antigen is the most immunogenic antigen in the complex Rh blood group system discovered in the year 1939. There is a lot of polymorphism in its phenotype due to genetic heterogeneity. Certain mutations and /or deletions lead to a weak phenotype defined by decreased density of antigen sites which require the use of anti human globulin for detection. The need for detection of the weak D antigen was to prevent alloimmunization by this blood if transfused to a D negative patient especially to women in child bearing age group. This contention is however, controversial and not proven beyond doubt. Moreover, the use of potent monoclonal D typing antisera detects low density of weak D antigens thus obviating the use of anti human globulin. We have assessed the incidence of Rh negative and weak D blood groups in the Garhwal region of Uttarakhand and reviewed the literature regarding the controversies in the clinical significance of weak D antigen.

Genotipagem RhD fetal não invasiva no acompanhamento de gestantes RhD negativo / Non invasive Fetal RHD Genotyping in management of pregnant RhD negative women

O acompanhamento de gestantes de fenótipo RhD negativo é baseado na premissa de que seus fetos podem estar em risco de desenvolver a doença hemolítica perinatal (DHPN) ou eritroblastose fetal, trazendo sérios riscos ao feto em decorrência de hemólise, com consequente anemia, hidropsia e, por vezes, óbito intrauterino. Procedimentos invasivos como amniocentese ou cordocentese podem ser utilizados para se inferir o fenótipo RhD fetal, entretanto, oferecem riscos ao feto e à gestante. Nos últimos anos, o conhecimento sobre a genética dos grupos sanguíneos e o desenvolvimento de técnicas de biologia molecular tem permitido a inferência de fenótipos de grupos sanguíneos a partir da detecção do material genômico. Inicialmente, a genotipagem do DNA fetal para o gene RhD era feita a partir de amostras de amniócitos ou de vilosidades coriônicas. No entanto, por tratar-se de testes invasivos, traziam risco ao feto e à gestante. A possibilidade de se obter DNA fetal a partir do plasma materno foi um grande avanço na prática clínica, por ser um procedimento não invasivo e, portanto, isento de risco. Esta revisão apresenta os princípios da técnica e os resultados de diferentes protocolos para genotipagem RhD fetal (publicados ao longo dos anos) e qual o seu propósito no acompanhamento das gestantes RhD negativo

The RhD negative pregnant women management has been based on the fact that their fetuses may be at risk of developing hemolytic diseases (DHPN) or erythroblastosis fetalis. This condition may bring serious risks to the fetus due to hemolysis, with consequent anemia and hydrops and sometimes, intrauterine death. Invasive procedures such as amniocentesis or cordocentesis may be performed to assess the fetal RhD phenotype, however, it offers risks to both fetus and pregnant woman. In recent years, the knowledge about the genetics of blood groups and the development of molecular biology techniques has allowed the inference of blood group phenotypes by the detection of genomic material. Initially, the fetal DNA genotyping for the RHD gene was performed from amniocytes or chorionic villi samples. Unfortunately, these invasive tests could bring risk to the fetus and the pregnant woman. However, the possibility of obtaining fetal DNA from maternal plasma has been a major advance in clinical practice, as being a non-invasive procedure and therefore not providing any risks. This review presents the principles of techniques and results of different protocols for fetal RHD genotyping (published over the years) and its goal on the management of RhD negative pregnant women

Human Sociogenetics

In three cities of Chile (Santiago, Valparaiso, Valdivia) the A allele and phenotype (ABO blood group) are more frequent in the higher socioeconomic strata (SES) and the O allele and phenotype are in the lower ones. This constitutes a structured sociogenetic cline (SGC). The B allele and phenotypes (B+AB) present a rather erratic or contradictory distribution among SES. This SGC was also found in England. The standard interpretation of the origin and maintenance of this SGC in Chile is founded on socio-ethno-historic-cultural and drift factors followed by socioeconomic assortative mating that has occurred since the origin of Chileans by the admixture of Europeans and Amerindians. This interpretation is insufficient to explain the coincidence of the cline in England and Chile, and for some findings in Chile. 1) The A and Rh(-) frequencies of the highest SES in Chile are significantly higher than those found in Europeans. 2) The B gene and phenotypes (with AB) behave differently and in contradiction to the socio-ethno-cultural-historical process. 3) There is a significant interaction of the SGC with gender in Chile and England. There is not at present a putative relationship between ABO and psycho-social factors that could account for this sociogenetic interaction. This SGC seems to be present in societies with a hierarchical organization in relation to power, prestige, ownership, income and life style, and when sampling includes the most extreme SES. It has not been found in two samples from Ireland and in a sample from Chile taken from a public hospital, probably because those variables and conditions were not ascertained.

Bases moleculares do sistema Rh e suas aplicações em obstetrícia e medicina transfusional / The molecular basis of RH system and its applications in obstetrics and transfusion medicine

O sistema Rh é o mais polimórfico e imunogênico de todos os sistemas de grupos sanguíneos. Atualmente mais de 49 antígenos foram identificados sendo cinco principais os antígenos D, C, c, E, e. O conhecimento das bases moleculares do sistema Rh desde a sua primeira clonagem há 17 anos possibilitou o entendimento tanto do mecanismo do fenótipo Rh negativo quanto das variantes dos antígenos RHD e RHCE. As deleções, rearranjos gênicos e as inserções são as principais mutações encontradas. Nos caucasianos, o mecanismo principal do fenótipo Rh negativo é a completa deleção do gene RHD, enquanto nos afrodescendentes é a presença do pseudogene RHDψ e do gene híbrido RHD-CE (4-7)-D. Os autores analisam a estrutura do complexo Rh nas hemácias, as bases moleculares do Sistema Rh, os mecanismos de negatividade RHD, além da Expressão fraca e parcial de D.

The Rh system is the most polymorphic and immunogenic for all blood group systems. Currently more than 49 antigens were identified with five major antigens D, C, c, E, e. Knowledge of the Rh system's molecular basis, since its first cloning 17 years ago, allowed to understand the mechanism of Rh-negative phenotype and the variants of antigens as RHD and RHCE. Deletions, gene rearrangements and insertions are the main mutations. In Caucasians the primary mechanism of Rh-negative phenotype is the complete RHD gene deletion, while in African descendants it is the presence of pseudogene and gene RHDψ hybrid RHD-CE (4-7)-D. The authors analyze the structure of the Rh complex in red cells, molecular basis of the Rh system, mechanisms of Negativity RHD and weak and incomplete expression of RHD.

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.

Primary anti-D Immunization by DEL Red Blood Cells / 대한진단검사의학회지

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.

Eficacia de la fenotipificación Rh / Effectiveness of Rh fenotipification

Basándonos en garantizar la calidad de los procedimientos del laboratorio pretransfusional, se evaluó la eficacia de la fenotipificación Rh en donantes y receptores. Para ello se comparó el número de pacientes sensibilizados transfundidos con glóbulos rojos desplasmatizados (GRD) con fenotipo Rh tomados al azar en relación con pacientes sensibilizados transfundidos con GRD isofenotipo, a fin de determinar si la implementación de las técnicas de fenotipificación Rh en dadores y receptores disminuye la sensibilización por este sistema. De los 113 anticuerpos hallados, 65 (57,52 por ciento) corresponden al sistema Rh y 48 (42,48 por ciento) a otros sistemas de antígenos eritrocitarios. Dentro de los 65 anticuerpos del sistema Rh, 56 (86,15 por ciento) pertenecen al primer grupo y 9 (13,85 por ciento) al segundo. A través de esta casuística demostramos que la implementación de la fenotipificación Rh disminuyó la sensibilización a este sistema.

In order to guarantee the quality of procedures in the pretransfusional laboratory, RH fenotipification efficiency was analised in donors and recipients. To do this, the number of sensitized patients transfusioned with red cells (GRD) with fenotype Rh taken at random was compared with patients transfusioned with fenotype GRD, in order to determine whether the implementation of RH fenotipification techniques in donors and recipients diminishes the sensitization by this system. Out of 113 antibodies found, 65 (57,52 per cent) correspond to Rh system and 48 (42,48 per cent) to other systems of antifungals red cells. Within the 65 antibodies of the Rh system, 56 (86,15 per cent) belong to the first group and 9 (13,85 per cent) to the second. Through these cases, we are showing that the implementation of fenotipification Rh diminished the sensitization to this system.