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.

Molecular genetic analysis of a patient with double population of red blood cells for RhD antigen / 中国输血杂志

【Objective】 To solve the difficulty of RhD blood group typing in a patient with double population(DP) of red blood cells for RhD antigen by serological and genotyping analysis. 【Methods】 Separation of the two populations of red blood cells of the patient was performed using capillary centrifugation method. ABO, RhD and RhCE typing, direct anti-human globulin test (DAT), irregular antibody screening, antibody identification and blood crossmatching of the patient were conducted using the standard serological methods. The hybrid Rhesus zygosity analysis of the RHD gene was performed by PCR-RFLP method. RHD and RHCE genotype of the patients were identified by PCR-SSP method. 【Results】 The patient was B type but with DP of red blood cells for RhD, Rhc and RhE antigens. DAT of the patient was positive and the alloanti-D was detected in serum. The RHD zygosity was D-/D- homozygote. PCR-SSP testing showed the RHD gene deletion (RHD * 01N. 01/01N.01 genotype) and Ccee of RHCE genotype in the patient, which was consistent with RHD zygosity analysis. 【Conclusion】 This is a special case with D-negative phenotype which was wrongly detected as D-positive type after D-positive red blood cells transfusion in emergency. When the DP of red cells for D antigen encountered like this case, the RhD typing can be accurately determined by using RHD genotyping analysis to provide strong evidence to the clinical blood transfusion.

Guía de manejo obstétrico y del recién nacido en madre aloinmunizada / Guidelines for the obstetric management of an alloimmune mother and her newborn / Guia de manejo obstétrico e neonatal em mães aloimunizadas

Resumen: La aloinmunización es una respuesta biológica frente a la exposición de antígenos no propios. La gestación, las transfusiones de hemocomponentes, los trasplantes de órganos sólidos y células hematopoyéticas, así como el consumo de drogas intravenosas exponen a las pacientes al desarrollo de aloanticuerpos antieritrocitarios. El hallazgo de los mismos debe cumplir con las instancias diagnósticas para identificar la probabilidad de estar asociados a enfermedad hemolítica feto neonatal (EHFN) y su oportuna derivación a policlínica de alto riesgo obstétrico (ARO) para su correcto seguimiento. Es fundamental que sean los laboratorios de inmunohematología de los servicios de hemoterapia y medicina transfusional los encargados de los estudios diagnósticos de aloinmunización eritrocitaria(1). En este sentido hemos elaborado esta guía con el objetivo de protocolizar de manera multidisciplinaria el manejo de las embarazadas aloinmunizadas y sus recién nacidos.

Abstract: Alloimmunization is the biological response to exposure to non-HLA antigens. Pregnancy, transfusion of blood components, solid organ and hematopoietic cell transplantation, as well as intravenous drug use expose patients to the development of anti-erythrocyte antibodies. When the latter are found, they must match diagnostic criteria to identify the potential association to hemolytic disease of the fetus and newborn (HDFN) and its timely referral to the high-risk obstetric risk polyclinic for due follow-up. It is of the essence for erythrocyte alloimmunization diagnostic tests to be carried out by the immunohematology laboratories of the Hemotherapy and Transfusional Medicine services. To that end, we have prepared these guidelines with the purpose of providing a multidisciplinary protocol for the handling of maternal alloimmunization and alloimmunization of the newborn.

Resumo: A aloimunização é uma resposta biológica à exposição a antígenos não próprios. A gravidez, as transfusões de hemocomponentes, os transplantes de órgãos sólidos e células hematopoiéticas, bem como o uso de drogas intravenosas expõem os pacientes ao desenvolvimento de anticorpos antieritrocitários. O achado destes deve obedecer a critérios diagnósticos para identificar a doença e a probabilidade de estarem associados a doença hemolítica feto neonatal (DHPN) e seu encaminhamento oportuno para uma unidade de alto risco obstétrico para acompanhamento adequado. É fundamental que os laboratórios de imuno-hematologia dos serviços de Hemoterapia e Medicina Transfusional se encarreguem dos estudos diagnósticos da aloimunização eritrocitária. Elaboramos este guia com o objetivo de estabelecer um protocolo multidisciplinar para o manejo de gestantes aloimunizadas e seus recém-nascidos.

Follow up of hemolytic changes in newborns induced by high titer IgG anti-D / 中国输血杂志

【Objective】 To analyze the results of maternal anti-D measured prenatally and serological tests of the newborn postnatally, so as to provide a basis for perinatal prevention and treatment of hemolytic disease of the newborn(HDN) in Rh negative pregnant women. 【Methods】 Irregular antibodies screening, antibody identification and titer determination were carried out for pregnant women prenatally. Blood group typing and 3 HDN tests were performed for the infant suspected as HDN. One-month follow up, since the delivery of the infant, concerning the changes in antibody titers, hemoglobin and bilirubin of the infant were monitored. 【Results】 The anti-D titer increased to 512 since 28 weeks in the parturient, 1 024 at 30 weeks of gestation, and decreased to 512 at the time of delivery. The masking effect on RhD antigen was observed after the birth of the child, with a sustained decrease in hemoglobin and a sharp increase in total and indirect bilirubin after 24 h of birth, which peaked within 48 h and gradually decreased thereafter.The antibody titers gradually decreased after birth. 【Conclusion】 Close monitoring of the changes of irregular antibody titer in Rh negative parturients is helpful for the prevention and early treatment of HDN, and hemoglobin changes in the newborn should be monitored immediately after birth, as well as symptomatic treatment to reduce the involvement of affected children.

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.

Hemolytic disease of newborn caused by multiple Rh antibodies

Hemolytic disease of Fetus and Newborn (HDFN) usually results due to natural occurring antibodies or alloimmunization in mother but the presence of multiple red cell antibodies increases the risk of development of significant HDFN. Here author reported a case of hemolytic disease of fetus and newborn in a preterm baby caused by multiple maternal antibodies. Direct Antiglobulin Test (DAT) on neonate blood sample was positive (3+) with monospecific DAT showed IgG type which was confirmed by heat elution. Antibody identification of eluate was done using commercial 11-cell panel by gel method showing specificity to anti-D and anti-C antibody which was differentiated from anti-G by sequential adsorption and elution studies. Neonate was treated with double volume exchange transfusion (DVET) using leucoreduced, irradiated O Rh D and C negative PRBC suspended in AB plasma and discharged 6th day in a stable condition. So, all pregnant women should be at least advised for ICT irrespective of Rh D negative status. If ICT is positive, they should be referred to higher center for proper Immunohematological work up, so that proper blood unit for DVET could be identified.

Prevalance of Rh Antigen Subgroups Among Blood Donors in a Tertiary Care Hospital in Rural Harayana

Rh system in today’s world is probably the most complex red cell antigen system in humans. The presence ofD antigen confers Rh positivity and vice versa. Two allelic antigen pairs, E/e and C/c are also found on the Rhprotein. The D antigen is the most immunogenic red cell antigen after A and B. As there is paucity of datadistribution of Rh antigen subgroup from the Indian literature, the study was conducted to know the prevalenceof Rh antigen subgroups in this part of the region and to determine the phenotype and most common genotypeof Rh antigen among the blood donors. The observational 1 year prospective study was conducted on blooddonors attending the blood bank in the Department of Pathology, MMIMSR, MMDU, Mullana.The studycomprised of blood donors of various age groups which included 90%(450) males and 10%(50) females. Anoverall Rh D positivity was seen in 88.4% of blood donors while 11.6% lacked the D antigen. The mostcommon Rh phenotype was ccDEE 26.6%. In conclusion, sensitization to clinically important blood groupantigens can be prevented through complete blood typing. All patients should be genotyped before the firstblood transfusion

Prevalencia de anticuerpos irregulares en donantes en un banco de sangre de Antioquia, 2016-2018 / Prevalence of irregular antibodies in donors of the blood bank in Antioquia, 2016-2018

Resumen Introducción: la aloinmunización se presenta entre 1,0-1,5 % de las personas expuestas a un antígeno externo o durante el embarazo, un trasplante o una transfusión. En los bancos de sangre se realiza la tamización de anticuerpos irregulares, diferentes a los del sistema ABO. Objetivo: estimar la prevalencia de anticuerpos irregulares en donantes de un banco de sangre de Medellín entre 2016 y 2018. Métodos: estudio transversal en 25 391 donantes. Se realizó control de sesgos de selección e información mediante aplicación de técnicas con excelente validez y control de calidad interno y externo, y detección mediante ID-card Liss/coombs en seis microtubos con anti-IgG y anti-C3d. El análisis se basó en medidas de resumen, frecuencias, chi cuadrado y prueba exacta de Fisher. Resultados: la edad promedio fue 34 años. La prevalencia de anticuerpos irregulares fue 0,30 %, siendo mayor anti-D con 0,071 %. No se hallaron diferencias según el sexo y el grupo etario; anti-D fue más frecuente en mujeres y donantes entre 31 y 40 años. Conclusión: se halló una alta prevalencia de anti-D y antiKell, que presentan gran importancia clínica después de los anticuerpos del sistema ABO. Esto evidencia la necesidad de esta prueba para reducir las reacciones post-transfusionales.

Abstract Introduction: Alloimmunization occurs between 1.0-1.5 % of people exposed to an external antigen or during pregnancy, a transplant or transfusion. In the blood banks, the screening of irregular antibodies different from those of the ABO system is performed. Objective: To estimate the prevalence of irregular antibodies in donors of a blood in Medellin 2016-2018. Methods: Cross-sectional study in 25 391 donors. Control of selection and information bias was carried out by applying techniques with excellent validity and internal and external quality control, and detection by ID-card Liss / coombs in six microtubes with anti-IgG and anti-C3d. The analysis was based on measures of summary, frequencies, chi-square and Fisher's exact. Results: The average age was 34 years. The prevalence of irregular antibodies was 0.30%, being higher anti-D with 0.071%. No statistical differences were found according to sex and age group; Anti-D was more frequent in women and donors aged 31-40 years. Conclusion: A high prevalence of Anti-D and Anti-Kell was found, which are of great clinical importance after the antibodies of the ABO system. This demonstrates the need for this test to reduce post-transfusion reactions.

Fetomaternal hemorrhage in RhD-negative pregnant women in Tianjin / 国际生物医学工程杂志

Objective To study and monitor the situation of femomaternal hemorrhage (FMH) in RhD-negative pregnant women in Tianjin, obtain the FMH data of such population, and analyze the relationship between FMH and age, blood type, gestational age, hemolytic disease of postpartum neonates, etc. Methods The FMH level was detected by flow cytometry with FITC-anti-HbF monoclonal antibody. The blood type was detected by blood serum method. The irregular antibody was identified by saline method and indirect anti-human ball method. The hemolysis of postpartum neonates was detected by three tests of hemolysis. Results The FMH volume of 86 RhD negative pregnant women was between 0 and 11.48 ml, with an average of 1.82 ml. There were 63.95％of pregnant women showed a volume of FMH<2.0 ml, 23.26％between 2 and 4 ml, 11.63％between 4.0 and 10.0 ml, and 1.16％>10 ml. The proportion of lower FMH in pregnant women≤30 years old was>11.71％higher than that in the pregnant women>30 years old, but the difference was no statistical significant. There was no significant difference in FMH of pregnant women with O, A, B and AB types. The proportion of higher FMH in pregnant women with compatible ABO blood type with her husband was 12.46％ lower than that of the heterozygous cases, but the difference was no statistical significant. The proportion of higher FMH in the pregnant women with 28 to 32 weeks gestational age was 14.55％ higher than that of ≤28 weeks and was 35.32％ higher than that of >32 weeks, and the differences were statistical significant. Three samples in the 86 samples were positive for anti-D antibody, and their three hemolytic test results were strongly positive with the anti-D titer from 1:2 to 1:32 and the FMH volume from 1.50 to 6.93 ml. The proportion of lower FMH in the 10 pregnant women without postpartum hemolysis was 70％ higher than that in 5 pregnant women with postpartum hemolysis, but the differences were not statistical significant. Conclusions The results suggest that monitoring FMH content by flow cytometry can reflect FMH in Rh-negative pregnant women. The studies on the relationship between FMH and age, blood type, pregnant time and hemolytic disease of postpartum neonates can provide basically experimental data for standard use of anti-D immunoglobulin in pregnant women.

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.

Anticuerpos anti-Fya con reacción anamnésica asociada con anti-D en transfusión intrauterina: reporte de un caso / Anti-Fya antibodies with anamnestic reaction associated to anti-D in intrauterine transfusion: A case report

Resumen ANTECEDENTES La enfermedad hemolítica perinatal ocurre después de una transfusión sanguínea, que sensibiliza con antígenos eritrocitarios, hemorragia materno-fetal durante el embarazo o al momento del parto. La incidencia de anticuerpos anti-D ha disminuido de 14 a 0.1% en las madres D-negativas. No existe una inmunoglobulina que evite o disminuya la aloinmunización por otros antígenos eritrocitarios durante el embarazo. La incompatibilidad del grupo sanguíneo Duffy es una causa común de enfermedad hemolítica perinatal. OBJETIVO Exponer el caso de una paciente con hijo con enfermedad hemolítica perinatal por anticuerpos anti-Fya y anti-D tratado con transfusión intrauterina. CASO CLÍNICO Paciente de 22 años, con antecedente de múltiples transfusiones sanguíneas y datos clínicos de síndrome anémico. En la semana 28 del embarazo fue valorada para aplicarle inmunoglobulina anti-D. Luego de aplicarle dos unidades de concentrado eritrocitario Rh negativo se observó incompatibilidad (++) en fase de antiglobulina humana (Coombs), por esto se realizó el escrutinio de anticuerpos irregulares en gel, que resultó positivo en células I y II (+++). Enseguida se inició el protocolo de identificación de anticuerpos irregulares con un panel de 11 células, que reportó aglutinación en las células 1, 2, 3, 5, 6, 7, 8 y 11, sin mostrar especificidad. El estudio de adsorción del anticuerpo anti-D mostró células de antígeno D+ con las que se estableció el diagnóstico de anticuerpos anti-Fya y anti-D. El embarazo finalizó mediante cesárea con el nacimiento de un varón con grupo y Rh O positivo, de 30.1 semanas de gestación (talla de 40 cm y peso de 2000 g) con hidrops fetal. Se le realizaron ciclos de reanimación e ingresó a la unidad de cuidados intensivos neonatales, sin tratamiento farmacológico, y después de una hora de vida extrauterina falleció. La madre se dio de alta del hospital 36 horas después del puerperio, sin complicaciones adicionales. CONCLUSIÓN Los anticuerpos antieritrocitarios anti-Fya, solos o en combinación con otros anticuerpos, provocan enfermedad hemolítica perinatal severa. El laboratorio de inmunohematología tiene participación importante en el diagnóstico, seguimiento y tratamiento de la enfermedad hemolítica perinatal.

Abstract BACKGROUND Hemolytic disease of the fetus and newborn occurs after alloimmunization with red blood cells antigens by blood transfusion, maternal-fetal hemorrhage during pregnancy or at delivery. Currently, the incidence of alloimmunization by anti-D antibody has been reduced from 14% to 0.1% of D-negative mothers, however, there is no immunoglobulin that prevents or decreases alloimmunization by other red blood cells antigens during pregnancy. The incompatibilities of the Duffy blood group are a common cause of hemolytic disease of the fetus and newborn. OBJECTIVE To present the case of a neonate with perinatal hemolytic disease secondary to anti-Fya and anti-D antibodies managed with intrauterine transfusion. CLINICAL CASE A 22-year-old patient with a history of multiple blood transfusions and clinical data of anemic syndrome. In the 28th week of pregnancy it was evaluated for the application of anti-D immunoglobulin. The blood bank was asked for two units of Rh negative erythrocyte concentrate. Incompatibility (++) in the human antiglobulin phase (Coombs) was observed, so the irregular antibody gel was screened, which was positive in cells I and II (+++). An identification protocol for irregular antibodies was initiated with a panel of 11 cells, which reported agglutination in cells 1, 2, 3, 5, 6, 7, 8 and 11, without specificity. The adsorption study of the anti-D antibody showed D + antigen cells. The diagnosis of anti-Fya and anti-D antibodies was established. The pregnant woman was terminated by caesarean section, from which a male with a group was born and Rh O positive, of 30.1 weeks of gestation (size of 40 cm and weight of 2000 g) with fetal hydrops. He underwent resuscitation cycles, entered the neonatal intensive care unit, without pharmacotherapy and died after one hour of extrauterine life. The mother withdrew 36 hours after the puerperium, without additional complications. CONCLUSION The antibodies anti-Fya alone or next to other alloantibodies produce severe hemolytic disease of the fetus and newborn. The laboratory of immunohematology in the blood bank is an essential tool in the diagnosis, monitoring and treatment of hemolytic disease of the fetus and newborn.

Prevalence of Weak D (Du) in Blood Donors in a Referral Teaching Hospital

Background: Rh D is the most important Blood Group antigen after ABO Blood group antigen for transfusion purpose. All negative blood units by routine methods must be tested to detect weak D using IAT method. When the test for D and Du is positive, the label should read Rh(D) Positive . When the test for D and Du is negative, the label should read Rh(D) Negative. Objective: To know the prevalence of weak D in the donor population. No study has been done in this part of the country earlier. It will help in the knowledge of weak D, which is very important for better patient care and prevent allo-immunzation in blood recipients. Materials and Methods: Blood samples were tested by ID Gel technique or by tube method with two anti D reagents - anti-D IgM monoclonal and blend of anti-D IgM&IgG. All negative samples were further tested for weak D in IAT phase by LISS/Coombs' gel card. Results: A total of 13043 samples were tested from January 2011 to December 2013. 12196 were Rh positive and 847 were Rh D negative. Weak D was positive in 8 samples. Conclusion: The study shows the prevalence of weak D as 0.07% in blood donors who were primarily from in and around Jalandhar in Punjab. These donors may have posed problem to the recipients of blood and blood product and their detection prevented them from alloimunisation.

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.

Neutralization of anti-venom IgY against main activities of Deinagkistrodon acutus venom / 中国药理学通报

Aim To evaluate the potency of anti-D. acutus venom IgY neutralizing the main activities of D. acutus venom.Methods After mixing the different a-mounts of IgY with snake venom and incubating togeth-er,the main activities of snake venom were assayed by biochemical methods.Results The in vitro assays in-dicated that anti-D.acutus venom IgY obviously neu-tralized the activities of PLA2 ,5′-nucleotidase,hyalu-ronidase,metalloprotease and serine proteinase (fi-brinogenase)in D.acutus venom.Mouse experiments showed that the ED50 value of IgY for mouse was 1 131.09 μg.Conclusion Anti-D.acutus venom IgY antibodies have good effects in neutralizing D.acutus venom without the toxicities themselves.

Analyzing Antenatal Test and Outcomes of Rh-negative Pregnant Women / 实用妇产科杂志

Objective:To study antenatal immunity test and outcomes of Rh-negative pregnant women.Methods:287cases(1.25％ in 22880 cases)of Rh-negative pregnant women delivered in Shanghai Sixth People hospital from January 1 st,2010 to December 31,2016 were retrospectively analyzed.Rh(D) blood group was identified by a micro column gel method,and the same as serum anti-D antibody.The Rh phenotype detection was done Serology method and the hemolytic disease of the newborn was used indirect antiglobulin test,serum free antibody test,absorption and elution test.Some Rh negative pregnant women were implemented autologous blood at 37 weeks of gestation.Results:Among 287 cases of Rh-negative,12 cases had anti-D antibody and the positive rate was 4.18％.Among the 12 cases of anti-D positive,their Rh phenotype were all ccdee and they all had history of childbearing.In 90 cases which had one history of birth,there were ten cases had anti-D antibody (11.11％),in 15 cases which had two history of birth,there were two cases had anti-D antibody(13.33％).287 Rh negative pregnant women had 290 child births,among which 8 newborn had neonatal hemolytic disease,the incidence rate was 2.75％ (8/290).In the 12 cases of anti-D positive,there were 1 case died because of fetal neonatal hemolytic disease and 7 cases were cured and 4 cases were normal with free of jaundice symptoms.There were 146 pregnant women implementation of autologous donation safely.According to the delivery way of childbirth,287 cases were divided by cesarean section and vaginal delivery,comparing postpartum haemorrhage amount with autologous donantion and unautologous donation,respectively,the result showed that they had no significant difference(P ＞0.05).Conclusions:It should be pay attention to the pregnant women who had childbearing history and whose Rh phenotype is ccee,which tend to produce anti-D immune antibody.However,this does not necessarily lead to hemolytic disease.Based on maternal and fetus conditions,autologous donation is safe and valuable in Rh-negative pregnant.

Anti-LW Showing Anti-D Mimicking Specificity: A Case Report / 대한수혈학회지

LW antigens are expressed in higher intensities in D-positive blood cells than D-negative cells, which can result in false identification of anti-D in pretransfusion testing. Although several cases of anti-LW have been reported abroad, to the best of our knowledge, none have been reported in Korea. Herein, we report a case of anti-LW in a 58 year-old RhD positive patient with non-Hodgkin's lymphoma with a positive direct Coombs test and a suspicion of the presence of passive anti-D antibodies because of a history of intravenous immunoglobulin administration. However, during a 5-month follow up, the antibody was confirmed as anti-LW on grounds that it showed weakened reaction in dithiothreitol treated cells and enforced reaction in cord O+ cells when compared to the results from antibody identification panel cells.

A Case of Primary Anti-D Alloimmunization by RHD (c.1227G>A) DEL Red Blood Cell Transfusion / 대한수혈학회지

The Rh blood group D antigen is the most immunogenic of all antigens, next to ABO antigens. Anti-D immunization is clinically important since it may cause clinical problems, such as severe hemolytic transfusion reactions and hemolytic disease of the newborn. DEL is an extremely weak D variant that cannot be detected by basic serologic typing and is typed as D-negative without the absorption-elution techniques and RHD genotyping. Of the DEL phenotype, RHD (c.1227G>A) allelic variant is the most common in Korea. The DEL phenotype has been considered to carry only a few D antigens to induce anti-D immunization, but a few cases have reported that this allelic variant is capable of inducing anti-D immunization in a D-negative recipient, for which it is clinical significant. Herein, we present a case of primary anti-D alloimmunization in a RhD negative patient after receiving RHD (c.1227G>A) DEL red cell transfusion identified by serological and molecular tests, including RHD genotyping.

Use of Staphylococcal Protein-A and Streptococcal Protein-G for Detection of Red Blood Cells (RBC) Antibodies and Comparison with Other Techniques.

Background: Detection of red blood cells antibodies is important for the diagnosis of autoimmune hemolytic anemia, hemolytic disease of newborn, pre-transfusion testing and other problems. The aim of this study was to use Staphylococcal protein A (SpA) and Streptococcal protein G (SpG) as reagents in immunological tests for detecting red blood cells (RBC) antibodies and to compare the method with other techniques. Study Design & Methods: Sera from 60 patients, comprising forty-four anti-D positive sera from pregnant women and 16 from healthy controls were, used for the study. The anti-globulin gel test and the standard Coombs’ test were used to determine RBC antibodies in these sera and the result were compared with that of protein A and protein G tests. Results: With various degree of agglutination all 4 techniques detected the presence of RBC antibodies (anti-D) in the sera from 44 pregnant women, and tested negative for the remaining 16 sera (from healthy controls). The sensitivity and the specificity of the 4 techniques was 100%. Conclusions: This preliminary study demonstrates that both SpA and SpG tests can be used for the detection of RBC antibodies and therefore requires more study and testing before they can become useful standard tests in transfusion medicine.

Red cell alloimmunization among antenatal women attending a tertiary care hospital in south India.

Red cell alloimmunization among antenatal women attending a tertiary care hospital in south India Jophy Varghese, Mary P. Chacko, Molly Rajaiah & Dolly Daniel Department of Transfusion Medicine & Immunohaematology, Christian Medical College & Hospital, Vellore, India Received December 13, 2011 Background & objectives: Detection of maternal alloimmunization against red cell antigens is vital in the management of haemolytic disease of the foetus and newborn (HDFN). This study was conducted to measure the presence of allosensitization to blood group antibodies in the antenatal women attending a tertiary care hospital and to observe the proportion of minor blood group antibodies to assess the benefit of screening for the same. Methods: All antenatal women registered in the hospital between January 2008 and January 2009, were screened for irregular antibodies using a commercial 3-cell antibody screening panel. Antibody identification was performed on samples found positive using a commercial 11 cell-panel. Results: Screening was performed on 5347 women, 339 (6.34%) of whom were Rh negative. Allosensitization was found in 79 women (1.48%; confidence interval 1.17 -1.84). In 29 of these 79 (37%) women the allo-antibodies could not be identified. In the remaining 50 women, 54 antibodies were characterized. A total of 40 clinically significant antibody specificities were identified among 36 women, of whom four were Rh(D) positive. Allosensitization with clinically significant antibodies was found in 9.43 per cent (confidence interval 6.55-13.06) Rh(D) negative and in 0.08 per cent (confidence interval .02-0.2) Rh(D) positive women. Anti D was the most frequent antibody found in 8.85 per cent Rh(D) negative women. The remaining clinically significant antibodies identified included anti-C, c, E, Jka, Jkb, M and S. In Rh(D) negative women, anti-D and antibodies of the Rh system contributed 83.3 and 94.4 per cent of clinically significant antibodies. However, in Rh(D) positive women, non-Rh antibodies comprised three out of four clinically significant antibodies. Interpretation & conclusions: The presence of alloimmunization in our study corroborated with data reported from India. The most frequent antibody was anti-D. However, a significant fraction was non-D. Alloimmunization among Rh(D) positive women though low as compared to Rh(D) negative women, included clinically significant antibodies, and most of these were non Rh.