夫妇ABO血型不合O型血孕妇免疫球蛋白G抗A/B效价与新生儿溶血病的相关性研究

Correlation between immunoglobulin G anti-A/B titer and neonatal hemolytic disease in pregnant women with O-type blood group from ABO blood group incompatibility couples

目的探讨夫妇ABO血型不合O型血孕妇免疫球蛋白(Ig)G抗A/B效价与新生儿溶血病(HDN)的相关性。方法选择2015年2月至2017年8月,于山东省血液中心血型参比研究室进行夫妇ABO血型不合产前血型抗体检查的104例O型Rh阳性血型孕妇为研究对象。孕妇年龄为(26.5±6.8)岁。根据孕妇的孕次,将其分为3组:首次妊娠组(n=31),2次妊娠组(n=35),≥3次妊娠组(n=38)。采用微柱凝胶法检测孕妇及其丈夫的ABO血型、孕妇的Rh血型及血清IgG抗A/B效价。对新生儿脐静脉血标本进行HDN 3项试验,包括红细胞直接抗人球蛋白试验、血清游离抗体试验和红细胞抗体释放试验。根据以上检测结果,分别计算孕妇血清IgG抗A/B阳性率和新生儿ABO-HDN发生率,并且分析孕妇孕次、血清IgG抗A/B效价、新生儿ABO-HDN发生率的相关性。采用方差分析比较3组孕妇的年龄、孕龄等的总体差异。采用χ2检验对3组孕妇血清IgG抗A/B阳性率,母体不同血清IgG抗A/B阳性率的新生儿ABO-HDN发生率进行比较。以上资料进一步两两比较,采用Bonferroni法调整检验水准。采用Spearman轶相关分析孕妇的孕次、血清IgG抗A/B效价、新生儿ABO-HDN发生率3者的相关性。本研究遵循的程序符合山东省血液中心医学伦理委员会制定的标准,经过该伦理委员会批准(批准文号:鲁血伦审字[2019]5号),并与所有受试者及其监护人签署临床研究知情同意书。结果①本研究104例孕妇的ABO血型均为O型,Rh血型均为Rh阳性。其丈夫的ABO血型包括,A型41例、B型48例和AB型15例。新生儿的ABO血型包括,A型45例,B型50例,O型9例。②本研究104例孕妇的血清中,IgG抗A/B总阳性率为65.4%(68/104)。初次、2次、≥3次妊娠组孕妇血清IgG抗A/B阳性率分别为45.2%(14/31)、65.7%(23/35)、81.6%(31/38),3组比较,总体差异有统计学意义(χ2=10.007,P=0.008)。其中,≥3次妊娠组孕妇血清IgG抗A/B阳性率最高,与首次妊娠组比较,差异有统计学意义(χ2=9.981,P=0.002),与2次妊娠组比较,差异无统计学意义(χ2=2.382,P=0.123)。③本研究104例新生儿中,ABO-HDN发生率为17.3%(18/104)。母体血清IgG抗A/B效价<64、=64、=128、≥256的新生儿ABO-HDN发生率分别为5.6%(2/36)、9.5%(2/21)、25.0%(5/20)和33.3%(9/27),4者比较,总体差异有统计学意义(χ2=10.345,P=0.018)。其中,母体血清IgG抗A/B效价≥256的新生儿ABO-HDN发生率最高,与母体血清IgG抗A/B效价<64的新生儿比较,差异有统计学意义(χ2=8.261,P=0.004),与母体血清IgG抗A/B效价=64、128的新生儿分别比较,差异均无统计学意义(P>0.05)。④本研究104例孕妇的孕次与其血清IgG抗A/B效价,与新生儿的ABO-HDN发生率;母体血清IgG抗A/B效价与新生儿的ABO-HDN发生率,均呈正相关关系(r=0.423、P=0.035, r=0.491、P=0.021, r=0.491、P=0.021)。结论夫妇ABO血型不合O型血多次妊娠孕妇的血清IgG抗A/B效价阳性率升高,新生儿发生ABO-HDN的风险较大。

Objective To investigate the correlation between immunoglobulin (Ig) G anti-A/B titer and neonatal hemolytic disease (HDN) in pregnant women with O blood group from ABO blood group incompatibility couples. Methods From February 2015 to August 2017, a total of 104 pregnant women with O-type and Rh-positive blood group who underwent prenatal blood group antibody test because of couples with ABO blood group incompatibility in the Blood Type Reference Laboratory of Blood Center Shandong Province were selected as subjects. The average age of pregnant women were (26.5±6.8) years. According to gravidity of these pregnant women, they were divided into 3 groups : first pregnancy group (n=31), twice pregnancy group (n=35) and≥3 times pregnancy group (n=38). Micro-column method were performed to detect the ABO blood group of the couples, the Rh blood group and titer of serum IgG anti-A/B of pregnant women. The umbilical vein blood samples of newborns were tested by direct anti-globulin test, free antibody test and antibody elution test. The positive rate of serum IgG anti-A/B in pregnant women, the incidence of ABO-HDN in newborns, as well as the correlation among gravidity, serum IgG anti-A/B titer and incidence of ABO-HDN were calculated, respectively. The age and gestational age of pregnant women among 3 groups were compared by variance analysis. The positive rate of serum IgG anti-A/B of pregnant women among 3 groups, and the incidence of ABO-HDN of newborns among different maternal serum IgG anti-A/B titer, were compared by chi-square test, respectively. And further pairwise comparison of above data used Bonferroni method to adjust the significance level. The correlation among the gravidity maternal, serum IgG anti-A/B titer, and neonatal ABO-HDN incidence were analyzed with Spearman rank correlation analysis. The procedures followed in this study were in accordance with the standards established by the Medical Ethics Committee of Blood Center Shandong Province, and this study was approved by the committee. All the subjects signed the informed consents for clinical trials. Results ① In this study, ABO blood group of all 104 pregnant women were O-type, and Rh blood group were positive. The ABO blood group of their husbands included 41 cases of A-type, 48 cases of B-type and 15 cases of AB-type. The ABO blood group of the newborns included 45 cases of A-type, 50 cases of B-type and 9 cases of O-type.② Among the 104 pregnant women in this study, the total positive rate of serum IgG anti-A/B was 65.4%(68/104). The positive rates of serum IgG anti-A/B of pregnant women in the first, twice,≥3 times pregnancy groups were 45.2%(14/31), 65.7%(23/35) and 81.6%(31/38), respectively. The overall difference was statistically significant among the 3 groups (χ2=10.007, P=0.008). The positive rate of serum IgG anti-A/B of pregnant women in the ≥3 times pregnancy group was higher than that of the first pregnancy group, and the difference was statistically significant (χ2=9.981, P=0.002). But there was no significant difference between the first group and ≥3 times pregnancy group (χ2=9.981, P=0.002).③ Among 104 newborns in this study, the incidence of ABO-HDN was 17.3%(18/104). The incidences of ABO-HDN in newborns of maternal serum IgG anti-A/B titer<64,=64,=128,≥256 were 5.6%(2/36), 9.5%(2/21), 25.0%(5/20) and 33.3%(9/27), respectively. And compared these four, the overall difference was statistically significant (χ2=10.345, P=0.018). The incidence of ABO-HDN in newborns of maternal serum IgG anti-A/B titer≥256 was the highest, which compared with newborns of maternal serum IgG anti-A/B titer <64, the difference was statistically significant (χ2=8.261, P=0.004), but compared with newborns of maternal serum IgG anti-A/B titer =64 and 128 respectively, the differences were not statistically significant (P>0.05).④ In this study, the gravidity of 104 pregnant women were positively correlated with their serum IgG anti-A/B titers, which were positively correlated with the incidences of ABO-HDN in their newborns, and the maternal serum IgG anti-A/B titers was positively correlated with the incidences of ABO-HDN in their newborns (r=0.423, P=0.035;r=0.473, P=0.031;r=0.491, P=0.021). Conclusions The serum IgG anti-A/B positive rate of O-type blood group pregnant women with multiple pregnancies from ABO blood group incompatibility couples increased, and the risk of ABO-HDN in their newborns was high.