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.

Use of prophylactic phototherapy for RhD neonatal disease in a referral service

Abstract Objective: This study aimed to describe the effect of prophylactic phototherapy in the treatment of infants with Neonatal Hemolytic Disease. Method: A retrospective cohort study was carried out with 199 RhD-positive infants, born to RhD-negative mothers, alloimmunized for RhD antigen, between January 2009 and December 2018. Results: The incidence of exchange transfusions in the study population was 9.5%, with a mean maximum bilirubin value of 11.3 mg % (± 4.3mg %). Bilirubin's maximum peak was achieved with a mean of 119.2 life hours (± 70.6h). Conclusions: The low incidence of exchange transfusion, the extended maximum bilirubin peak for later ages, and the low mean of the maximum bilirubin values may indicate a positive effect of prophylactic phototherapy in the treatment of this disease. Further studies must be carried out to confirm these findings.

Hemolytic disease of newborn caused by unexpected antibodies: a retrospective study of 287 cases / 中国输血杂志

【Objective】 To observe the distribution of non-ABO-HDN and its clinical relevance, so as to provide reference for clinical diagnosis and treatment. 【Methods】 A total of 287 cases of non-ABO-HDN recorded during January 2012 to August 2022 were enrolled and tested in our laboratory. The correlation between maternal history of blood transfusion, pregnancy, unexpected antibody titers, gender, ABO-HDN and transfusion therapy was analyzed by chi-square test. 【Results】 All 287 cases of non-ABO-HDN involved 13 kinds of unexpected antibodies of 6 blood group systems. Rh-HDN accounted for 96.17% (276/287), and anti-D-HDN accounted for 47.04% (135/287). The proportion of non-ABO-HDN patients without ABO-HDN requiring exchange/transfusion was significantly higher than that of non-ABO-HDN patients with ABO-HDN(P8) was significantly higher than that in the low titer group (≤8) (P<0.05). There was no significant difference in gender, mother′s history of blood transfusion, pregnancy and whether or not to exchange/transfusion (severity of illness). 【Conclusion】 Understanding the characteristics of non-ABO-HDN and the specific distribution of unexpected antibodies, the correlation between various factors and diseases and their clinical significance are conductive to timely taking necessary intervention measures and reducing the risk of complications.

Analysis of ABO system hemolytic disease of the fetus and newborn in 1 040 cases and its influencing factors / 中国输血杂志

【Objective】 To analyze ABO system hemolytic disease of the fetus and newborn (HDFN) and its influencing factors in Obstetrics Department of our hospital. 【Methods】 The blood samples of 1 040 neonates and their mothers in the obstetric department of our hospital were retrospectively analyzed from September 2022 to January 2023, including ABO and RhD blood group of the neonates and mothers, as well as 3 tests of HDFN, Hb, total bilirubin (TBIL) and indirect bilirubin(IBIL) of the neonates. Relevant clinical data of the neonates and mothers were collected, including maternal and neonatal age, neonatal sex, maternal pregnancy history, gestational age and delivery mode, and their influences on ABO-HDFN were analyzed. 【Results】 Among 1 040 HDFN samples, 298 were ABO incompatibility, among which 113 were HDFN positive, with a positive rate of 37.9% (113/298); the positive rate of HDFN in neonates born to mothers with type O was significantly higher than that in neonates born to mothers with type A and B (71.4% vs 8.2%, P<0.05); the positive rate of HDFN in neonates with antigen-A incompatibility was significantly higher than that in neonates with antigen-B incompatibility (48.7%vs 26.7%, P<0.05), which was the highest in neonates with O-A incompatibility [83.6% (61/73)], followed by O-B incompatibility [58.2% (39/67)]. There was no significant difference in Hb and bilirubin among the other groups except for the difference of Hb between the O-A incompatibility HDFN positive group and the HDFN negative group [(145.0±16.0) vs(153.4±13.2), P<0.05)]. The levels of Hb, TBIL and IBIL in the "direct antiglobulin test+ indirect antiglobulin test+release test+" group were significantly different from those in the HDFN negative group[(144.9±21.6) vs (153.3±13.2), P <0.05; (36.9±11.8) vs (29.6±6.1), P<0.05; (30.6±12.7) vs (23.0±6.9), P<0.05, respectively]. Logistic regression analysis showed that maternal delivery frequency, mother-neonate incompatible antigen and maternal blood type were independent risk factors for HDFN. 【Conclusion】 ABO-HDFN occurred mainly in neonates born to O-type mothers, and the positive rate was the highest in neonates with O-A incompatibility. The severity of HDFN had little relationship with the mother-neonate blood type, but had relationship with the result of 3 tests of HDFN. Maternal delivery frequency, mother-neonate incompatible antigen and maternal blood type were independent risk factors for HDFN.

Blood immunological analysis of hemolytic disease of the fetus and newborn caused by Rh deletion type D-- / 中国输血杂志

【Objective】 To analyze the causes of a case of hemolytic disease of the fetus and newborn (HDFN),and investigate the genetic background of maternal Rh deletion D--formation. 【Methods】 Blood samples of maternal and fetus were collected, and ABO blood typing, Rh blood typing, antibody screening and identification test were performed to explore the blood group serological characteristics of Rh deletion type D--, and Rh gene sequence was performed on parturient. 【Results】 The maternal blood group was identified to be O type, D--, and the anti-Hr0 antibody against Rh high-frequency antigen was suspected to be caused by multiple pregnancies which passes through the placental barrier and enable fetus to obtain anti Hr0 antibody, leading to HDFN, with genetic testing result as RH RHCE* Ce/RHCE* Ce. 【Conclusion】 In-depth research on the formation mechanism of Rh D-- in parturient should be conducted to provide clinical value for HDFN blood exchange treatment and blood transfusion in special blood group population.

Severity and multiple indexes of jaundice in ABO hemolytic disease of fetus and newborn: a comparative analysis / 中国输血杂志

【Objective】 To explore the correlation between the severity of jaundice in ABO hemolytic disease of fetus and newborn (HDFN) and multiple indicators, in order to achieve accurate diagnosis, timely treatment, and prevent complications in ABO HDFN. 【Methods】 From March 2020 to February 2023, a total of 283 ABO HDFN in our hospital were classified into mild, moderate and severe groups based on total bilirubin levels. The differences in basic data, relevant laboratory indicators and the agglutination intensity of the three hemolysis tests were analyzed and compared. 【Results】 In the severe group, 75% (12/16) were male infants, which was significantly higher compared to 37.5% (57/152) in the moderate group (P<0.05). The values (mg/dL) of transcutaneous jaundice in the mild, moderate and severe groups were:forehead 9.88±1.93 vs 12.34±2.01 vs 15.56±2.69, face 10.25±2.27 vs 13.28±2.32 vs 15.99±2.86, chest 9.67±2.16 vs 12.51±2.11 vs 15.33±2.36, respectively. The values(µmol/L) of direct bilirubin (DBIL) and indirect bilirubin (IBIL) were 9.87±2.06 vs 11.90±2.59 vs 16.12±4.73, and 159.36±37.55 vs 252.98±30.52 vs 353.76±55.68, respectively, and the differences between the mild, moderate and severe groups were statistically significant (all P<0.05) . The creatine kinase (CK) values(unit/L) of mild, moderate and severe groups were 664.15±498.26 vs 500.51 ±451.63 vs 402.63±224.27, and the difference between the mild group and the other two groups was statistically significant (P<0.05) . There was no statistically significant difference in the agglutination intensity of the three hemolysis tests among HDFN with different severity of jaundice. 【Conclusion】 Clinicians can predict the severity of jaundice in light of gender, transcutaneous jaundice, DBIL, IBIL and CK of ABO HDFN, for further and graded treatment, so as to avoid the sequelae or even life-threatening consequences caused by ABO HDFN.

Value of three hemolysis tests and carboxyhemoglobin in the diagnosis of hemolytic disease of the fetus and newborn / 中国输血杂志

【Objective】 To investigate the value of three hemolysis tests and carboxyhemoglobin (COHb) level in the diagnosis of hemolytic disease of the fetus and newborn (HDFN). 【Methods】 From January 1, 2019 to December 31, 2022, the neonates hospitalized in the Department of Neonatology of Hebei Provincial Children's Hospital with suspected hemolytic disease who had serological testing were retrospectively enrolled in the study. They were distributed into HDFN group and non-HDFN group according to the final diagnosis. Their clinical and laboratory data were collected and analyzed, and the COHb level was detected by blood gas analyzer. 【Results】 A total of 378 neonates with HDFN and 217 neonates without HDFN were included in the study. Most of the neonates in HDFN group were full-term infants (348/378, 92.1%), with median gestational age of 39.1 (38.3, 40.0) weeks. Three hundred and fifty-four cases (354/378, 93.7%) were ABO-HDFN and the rest were Rh HDFN. There were significant differences in the level of serum total bilirubin, hemoglobin, COHb and reticulocyte percentage at admission between the two groups(P<0.05). The positive rate of three hemolysis tests in HDFN group decreased with the increase of the days after birth. The highest positive rate (more than 80%) was observed within 2 days after birth. Correlation analysis showed a negative relationship between the COHb level and the age (rs = -0.434, P<0.001) . Among the three hemolysis tests in HDFN group, the positive rate of antibody release test was the highest (69.0%), followed by the free antibody test (55.6%) and the direct antiglobulin test (DAT) (36.0%). Receiver operating characteristic (ROC) curve analysis showed that the optimal cut-off value of COHb was 1.15%.The sensitivity of COHb ≥ 1.15% was 51.8%, higher than single DAT (36.0%) . The diagnosis effectiveness of three hemolysis tests couldn't be improved when combined with COHb detection(Z = -0.727, P>0.05) . 【Conclusion】 The three hemolysis tests are important in the diagnosis of HDFN, among which the antibody release test has the highest sensitivity. COHb has certain value for the diagnosis of HDFN, but joint testing cannot improve the diagnosis effectiveness of three hemolysis tests. Hemolysis tests and/or COHb detection should be conducted for neonates at risk of hemolysis as early as possible after birth.

Effect of IgG anti AB antibody on the correlation analysis between the titer of blood group antibodies and total bilirubin in serum of newborns with hemolytic disease / 中国输血杂志

【Objective】 To investigate the correlation between the titer of anti-A or anti-B antibodies before and after the absorption of IgG anti-AB antibodies in the serum of type O mothers with ABO hemolytic disease of fetus and newborn (ABO-HDFN) and the total bilirubin in the serum of the children. 【Methods】 Serum samples from 119 children diagnosed with ABO-HDFN and their mothers sent to the Beijing Red Cross Blood Center from January to December 2020 were selected, and clinical data of the children were collected. Three hemolysis tests and serum total bilirubin (TBIL) determination were conducted on the children. IgG anti-A or anti-B antibody titers were tested before and after the mother′s serum absorbed IgG anti-AB antibodies. Statistical analysis was conducted on the IgG antibody titers and the TBIL results of the children. The differences in TBIL results corresponding to different IgG antibody titers were compared. The Spearman test was used to analyze the correlation between the IgG anti-A or -B antibody titers and TBIL results before and after the absorption of IgG anti-AB antibodies. 【Results】 There were differences in the TBIL results corresponding to IgG anti-A or anti-B titers at different levels in the serum of type O mothers after absorption by IgG anti-AB antibodies (F=8.401, 19.622, P0.05). The IgG anti-A or anti-B titers of maternal serum absorbed by IgG anti-AB antibodies were positively correlated with neonatal TBIL results (r=0.487, 0.629, P<0.05). 【Conclusion】 There is a positive correlation between the titer of IgG anti-A or anti-B antibodies in the serum of type O mothers after absorbing IgG anti-AB antibodies and the TBIL results of ABO-HDFN children. The trend of increased total bilirubin in newborn serum ban be accurately predicted by detecting the titer level of IgG anti-A or anti-B antibodies in the serum of mothers after absorbing IgG anti-AB antibodies.

Retrospective Analysis of Irregular Antibodies Causing Hemolytic Disease of the Fetus and Newborn in Jiangxi Province / 中国实验血液学杂志

OBJECTIVE@#To analyze the characteristics of antibody-specific distribution, laboratory detection results of hemolytic disease of the fetus and neonatal(HDFN) caused by irregular blood group antibodies other than ABO, and its correlation with the clinical situation.@*METHODS@#The non-ABO-HDFN cases in our hospital from October 2012 to December 2021 were selected as the research objects, and the cases diagnosed with ABO-HDFN in the same period were randomly selected as the control group, and the data of antibody specific distribution, total bilirubin, direct antibodies, maternal history, age of the children, the presence or absence of combined ABO-HDFN, and whether to exchange/transfuse blood were retrospectively analyzed. The characteristics of non-ABO-HDFN in Jiangxi province were analyzed.@*RESULTS@#The detection rate of non-ABO-HDFN in Jiangxi province increased. Among 187 non ABO-HDFN cases, the highest percentage of Rh-HDFN was detected (94.6%). Compared with the control group of ABO-HDFN, the non-ABO-HDFN had higher mean integral value of direct antibody, higher peak total bilirubin, and longer duration. Anti-M-HDFN may have severe disease but the direct antibody weak positive/negative, it was easy missed in clinical and delayed the treatment. There is no correlation between the specificity of irregular antibodies, the sex of the child, the mother's previous childbirth history, the presence or absence of combined ABO-HDFN and the need for blood exchange/transfusion(P>0.05).@*CONCLUSION@#The irregular antibodies of causing non ABO-HDFN in Jiangxi area are mainly Rh blood group system, followed by MNS blood group system. Understanding the characteristics of HDFN disease, serological features and the correlation with clinical indexes will help to detect and treat non ABO-HDFN in time and reduce the risk of complications.

Clinical analysis of severe hyperbilirubinemia caused by hemolytic disease of the newborn and G6PD deficiency / 中国新生儿科杂志

Objective:To study the clinical characteristics and differences of severe hyperbilirubinemia caused by hemolytic disease of the newborn (HDN) and glucose-6-phosphate dehydrogenase (G6PD) deficiency.Methods:From January 2014 to December 2021, newborns (gestational age ≥ 35 weeks and postnatal age ≤ 28 d) admitted to the Department of Neonatology of Hunan Children's Hospital with severe hyperbilirubinemia caused by HDN or G6PD deficiency were retrospectively analyzed. According to the etiology of hyperbilirubinemia, they were assigned into HDN group and G6PD deficiency group. The general conditions, clinical manifestations, laboratory results, treatment and prognosis of the two groups were compared using SPSS 26.0 software.Results:A total of 532 cases were in the HDN group and 413 cases in the G6PD deficiency group. The HDN group reached peak hyperbilirubinemia earlier than the G6PD deficiency group [3(2,5) d vs. 6(4,8)d, P<0.05]. The HDN group had lower peak value of total serum bilirubin [379.5(345.6,426.7) μmol/L vs. 486.4 (413.5,577.4) μmol/L] and lower incidence of anemia [37.4% (199/532) vs. 55.0% (227/413)]than the G6PD deficiency group.The incidence of anemia with elevated reticulocyte percent(Ret%) in the HDN group was higher than the G6PD deficiency group[66.3%(132/199) vs. 5.7%(13/227), P<0.05]. Compared with the G6PD deficiency group, the incidences of exchange transfusion and repeated (≥2 times) exchange transfusion, acute bilirubin encephalopathy(ABE) and the mortality rate after withdrawal of treatment in the HDN group were significantly lower ( P<0.05). Conclusions:Neonatal severe hyperbilirubinemia caused by HDN has early onset. G6PD deficiency caused hyperbilirubinemia has higher incidences of anemia, more severe jaundice and ABE, without increased Ret%.

Relationship of blood types and ABO hemolytic disease of the newborn / 中国新生儿科杂志

Objective:To compare the differences of hemolysis between OA and OB blood type in ABO hemolytic disease of the newborn (ABO-HDN), to study the clinical features of ABO hemolytic disease in different type.Methods:From January 2015 to June 2020, full-term ABO-HDN neonates born to blood type O mothers admitted to our hospital were enrolled in this retrospective study. The neonates were assigned into OA group and OB group. SPSS 25.0 software were used to analyze the clinical data.Results:A total of 755 cases were enrolled, including 364 cases of OA group and 391 cases of OB group. On hour-specific bilirubin nomogram, no significant differences existed in high intermediate risk zone between the two groups ( P>0.05). In the low risk zone and the low intermediate risk zone, the proportion of newborns in OA group was higher than the OB group ( P<0.05). In the high risk zone, the proportion of newborns in the OB group was higher than the OA group ( P<0.05). The age of admission of the OB group was younger than the OA group ( P<0.05). The incidences of immunoglobulin usage and blood transfusion in the OB group were higher than the OA group ( P<0.05). No significant differences existed between the two groups in Coombs? test, antibody elution test, free antibody test, platelet count, reticulocyte percentage, the onset time of jaundice, the median serum total bilirubin level, the average hemoglobin level and the incidence of anemia on admission ( P>0.05). No significant differences existed in the incidence of exchange transfusion, the duration of phototherapy and hospitalization between the two groups ( P>0.05). Conclusions:Compared with OA incompatibility, newborns with OB incompatibility have higher incidences of hyperbilirubinemia, blood transfusion and younger age of admission. However, the two groups have similar rate of exchange transfusion and phototherapy and hospitalization duration.

Genetic diagnosis of MN and ABO hemolytic disease of the newborn complicated with pyruvate kinase deficiency: a case report and literature review / 中国输血杂志

【Objective】 To investigate the clinical and genetic characteristics of hemolytic disease of the newborn(HDN) induced by anti-M complicated with pyruvate kinase deficiency (PKD) disease. 【Methods】 The clinical data of a pregnant woman with unexplained adverse pregnancy outcome in the third trimester were retrospectively analyzed, and neonate anemia status and blood transfusion were followed up. With informed consent, peripheral blood of the neonate and her parents were collected for serological tests and disease-related gene target sequence capture and sequencing. The clinical and genetic characteristics of HDN induced by anti-M or PKD were reviewed. 【Results】 The neonate presented severe anemia, hepatosplenomegalism, hyperbilirubinemia at birth, and was confirmed MN combined with ABO neonatal hemolysis by serological tests. The neonate recovered by receiving blood exchange and phototherapy treatments, but he needed to receive blood transfusion each month because of hemolytic anemia. Genetic analysis showed that the neonate had compound heterozygous mutations (c. 1096C>T; c. 941T>C) of PKLR gene inherited from her parents. 【Conclusion】 The clinical manifestations of PKD are similar to that of HDN caused by anti-M in the early stage of the disease. Clinicians should exclude the metabolic diseases of red blood cells when diagnosing severe HDN caused by anti-M.

Pregnancy outcome with Rh blood group alloantibody: analysis of 71 pregnant women / 中国输血杂志

【Objective】 To retrospectively investigate the antibody distribution and pregnancy outcome of pregnant women with Rh alloantibody in Guangzhou, and summarize the prevalence, diagnosis and treatment experience of hemolytic disease of newborn (HDN) caused by Rh alloantibody, so as to provide data for the prevention, diagnosis and treatment of Rh-HDN. 【Methods】 A total of 17 345 pregnant women in Guangzhou from January 2014 to December 2020 were selected for irregular antibody screening test.Those with Rh antibody were followed up for delivery, and the clinical and laboratory examination results of pregnant women and newborns were analyzed. 【Results】 A total of 71 cases (0.41%, 17/17 345) with Rh alloantibodies were detected.Among them, anti-D, anti-E, anti-Ec, anti-C, anti-Ce, anti-c, anti-e accounted for 26.76% (19/71), 46.48% (33/71), 9.86% (7/71), 7.04% (5/71), 5.63% (4/71), 2.82% (2/71) and 1.41% (1/71), respectively.Among the 71 pregnant women, 34 gave birth to children with HDN, with the total prevalence rate of 47.89%, among whom 100%, 78.94% and 42.42% were anti-c, anti-D and anti-E, respectively.And 71.43% (5/7) of the children who underwent transfusion were with anti-D.Although the yield rate of anti-E was the highest, it involved low morbidity and mild symptoms, which preferred to occur in the first fetus.No significant difference in gestational age, birth weight and the occurrence time of jaundice was notice between the anti-D group and anti-E group, but the total bilirubin of the anti-E group was lower while the Hb level were higher than those of the anti-D group (P<0.05). Two children died, and others were cured by phototherapy, albumin, IVIG and blood transfusion. 【Conclusion】 The publicity of Rh-HDN for early prevention and treatment should be strengthened to improve the cure rate and the prognosis.

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.

Prevención de enfermedad hemolítica perinatal severa con inmunoglobulina intravenosa en paciente altamente sensibilizada / Prevention of severe perinatal hemolytic disease with intravenous immunoglobulin in a highly sensitized patient

RESUMEN La enfermedad hemolítica perinatal es infrecuente hoy por la prevención que de ella se hace. Sin embargo, existen casos de madres altamente sensibilizadas que desean tener un hijo, lo que obliga a que ese embarazo deseado sea controlado de manera especial y sometido a procedimientos invasivos no exentos de morbimortalidad fetal. El uso prenatal de inmunoglobulina humana en la madre puede representar una alternativa terapéutica. Se presenta un caso en que su uso impidió el desarrollo de enfermedad intrauterina y favoreció la buena evolución neonatal a pesar de que el pronóstico inicial era muy adverso.

ABSTRACT Perinatal Hemolytic Disease is uncommon today due to its prevention. However, there are cases of highly sensitized mothers who wish to have a child, that forces this desired pregnancy to be controlled in a special way and be subjected to invasive procedures not exempt from fetal morbidity and mortality. Prenatal use of human inmunoglobulin in the mother may represent a therapeutic alternative. We present a case in which its use prevented the development of intrauterine disease and favored a good neonatal evolution despite the fact that the initial prognosis was very adverse.

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.

Blood group serological testings of ABO hemolytic disease of newborn: A retrospective analysis / 中国输血杂志

【Objective】 To retrospectively analyze blood samples of suspected hemolytic disease of newborn(HDN)mothers and infants, and detect hemolytic disease caused by irregular antibodies, so as to provide help for the clinical diagnosis and treatment of HDN. 【Methods】 632 suspected HDN samples from Obstetrics and Pediatrics Department of our hospital from January 2016 to October 2018 were collected, and serologically detected by microcolumn gel technique, as well as DAT, free antibody test and antibody release test. 【Results】 Among 632 samples, 306 were HDN positive, with a positive rate at 48.4%, 64 suspected HDN, accounting for 10.1%, and 262 non confirmed HDN, accounting for 41.5%. 180 samples were type A, among which 145 were HDN positive, with a positive rate at 80.56%; 233 were type B, among which 157 were HDN positive, wiht a positive rate at 67.38%; 210 were type O, among which 1 was HDN positive, with a positive rate at 0.48%; 9 were type AB, among which 3 were HDN positive, with a positive rate at 33.33%. The positive rates of HDN differed by blood types (P<0.05). In 632 suspected HDN samples, 9 were with irregular antibody + immune anti-A, and 136 with solo immune anti-A; 10 were with irregular antibody + immune anti-B, and 170 with solo immune anti-B; 1 was with irregular antibody + immune anti-A and anti-B, and 2 with immune anti-A and anti-B; 4 HDN cases were caused by irregular antibody, while anti-S and anti-E cconstituted 2 and 2 cases, respectively. 【Conclusion】 ABO HDN is more common and attracts more attention in clinical practice than HDN scaused by other group systems, which were rare and easy to be ignored, but also may cause moderate and severe HDN. even severe anemia, edema and stillbirth of fetus. Therefore, it is necessary to carry out irregular antibody screening during pregnancy so as to achieve early detection and treatment.

Crossmatch incompatibility in infants caused by intravenous immunoglobulin-produced IgG antibody: Six cases / 中国输血杂志

【Objective】 To analyze the effect of intravenous immunoglobulin(IVIG)-produced IgG antibody on the crossmatch incompatibility of neonates. 【Methods】 Blood type grouping, antibody screening, crossmatch, direct anti-globulin test, elution test, indirect antiglobulin test, and IVIG titer determination were conducted by microcolumn gel method. 【Results】 IgG anti-A were detected out in the elution test and free antibody test of 6 infants, and the titer of IgG anti-A contained in IVIG was 256, which led to the crossmatch incompatibility between infants and donors with the same type. 【Conclusion】 The hemolysis and crossmatch incompatibility in newborns, born to ABO-compatible mothers, may occur due to the IVIG-induced IgG antibodies. The O-type washed red blood cells should be selected for transfusion.

Correlation between irregular antibody profiles of non-Rh blood group system and incidence of hemolytic disease of the newborn, 53 cases / 中国输血杂志

【Objective】 To retrospectively analyze the irregular antibodies in 6 blood group systems other than the Rh blood group system in 53 pregnant women and analyze its correlation with the occurrence of hemolytic disease of the newborn(HDN). 【Methods】 19 473 pregnant women were screened for irregular antibodies by microgel detection technology combined with anti-human globulin (IgG+ C3d), and the positive samples screened out were further confirmed to understand the types and titers of irregular antibodies. Irregular antibody type determination experiment: IgG type irregular antibody titer was determined after mercaptoethanol (2-Me) inactivated the serum of the irregular antibody positive specimen, and then IgG and IgM type were determined by comparing the titer levels of irregular antibody. Three hemolysis tests and total bilirubin tests were performed on umbilical cord blood during delivery to analyze the level of jaundice and the occurrence of HDN. 【Results】 53 cases of irregular antibodies other than the Rh blood group system were detected in 19 473 pregnant women, with a positive rate of 0.27%, mainly MNS and Lewis blood group system.The incidence of HDN was 39.6% (21/53). There were 27 cases of IgM, 7 IgG, and 19 IgM + IgG. Comparison of total bilirubin detection between the low titer group (≤8) and the high titer group (>8) : the latter was significantly higher than the former (P<0.05); IgG antibody subtypes: IgG1 of the latter significantly increased (P<0.05), and so was IgG3 in former (P<0.05). There was a significant positive correlation between IgG1, IgG3 and total bilirubin. The area under the curve of IgG1+ IgG3 for HDN diagnosis, the sensitivity and specificity were 0.953, 0.900, and 0.967, respectively. 【Conclusion】 Other than Rh blood group system, irregular antibodies are mainly distributed in MNS and Lewis blood group system. The incidence of HDN is higher in Kell, Duffy and Kidd blood group systems after producing irregular antibodies. Non-antibody types are mostly IgM type or IgM + IgG mixed, and the incidence of HDN is not high; Patients with poor maternal history, either high or low titer, can be classified into IgG1 and IgG3 in early stages, and those with Abnormal results should be included into the perinatal management of high-risk women with regular checking.

Correlation between Rh blood group antigen profiles in pregnant women and hemolytic disease of the newborn in Qingdao area: A retrospective study / 中国输血杂志

【Objective】 To investigate the profile of Rh blood group antigen in pregnant women and hemolytic disease of the newborn (HDN) in Qingdao area. 【Methods】 10 597 pregnant women admitted in our hospital during October 2016 to February 2020 were selected and the ABO, Rh blood group system antigen (D, C, c, E, e) and the irregular antibody were detected, and positive antibody was further identified. The irregular antibody of Rh blood group in pregnant women was statistically analyzed according to the history of blood transfusion and pregnancy. Twelve HDN cases were studied, and the results of ABO, Rh blood group antigen and irregular antibody, antibody property identification, HDN test and blood routine test were retrospectively analyzed. 【Results】 Among 10 513 cases of Rh-positive pregnant women, the common phenotype was CCee>CcEe>Ccee>ccEE>ccEe; among 84 cases of Rh-negative pregnant wome, the common phenotype was ccee>Ccee> CCee> ccEE>ccEe. The positive rate of irregular antibody was 1.06% (112/10597) in 10 597 pregnant women, of which the Rh antibody was the highest, rated at 56.25% (63/112). For 64 pregnant women with positive antibodies, antibodies against Rh system were different from those against other systems when stratified by the history of blood transfusion (P<0.05) and pregnancy (P<0.05). Twelve neonates were diagnosed with Rh-HDN, with IgG anti-E in 6 cases, IgG anti-D 3, IgG anti-cE 1, IgG anti-C 1and IgG anti-c 1. Among them, 3 were seriously ill and treated with blood exchange. 【Conclusion】 As two-child policy was implemented, the incidence of Rh HDN had increased. ABO, RhD, C, c, E and e matched transfusion should be administered for women at childbearing age. Meanwhile, clinical termination of delivery was recommended for pregnant women, who probably develop Rh-HDN and are with critical situation. Rh phenotype matched fresh blood should be prepared, which has great clinical significance for rescuing newborns.