RhD阴性个体遗传多态性与抗-D同种免疫关系研究
发布时间：2018-07-26 07:19
【摘要】：目的通过对血清学试验确认Rh D阴性个体的RHD基因型检测,研究不同类型遗传学背景Rh D阴性个体与抗-D同种免疫的关系。重点探讨各种Rh D变异型个体的献血及输血策略,为稀有血液的合理应用提供理论依据。方法1、对2011年3月至2013年6月医院送检的Rh阴性个体,通过询问输血史和妊娠史排除没有免疫史的个体。采用盐水试管法和间接抗人球试验(IAT)排除血清学试验常规试验阳性的个体。对孕妇的流产史及生育史进行分类,研究流产史及生育史对抗-D产生的影响。2、采用微柱凝胶法筛查抗体,并通过谱细胞进行抗体鉴定,确认抗体特异性。检测这些样本的Rh表型,包括D、C、c、E和e抗原。3、采用序列特异性引物PCR(PCR-SSP)技术及DNA序列分析技术,分析样本的RHD基因。人类红细胞RHD阴性鉴定基因检测试剂盒可检测的基因型包括Rh D阳性、Rh D阴性、Rh D-CE(2-9)-D、DVa(Hus)、DVI III型、弱D15型和DEL RHD1227A。结果1、本研究观察有免疫史Rh D阴性个体共计336例。其中22例有输血史的个体均为医院在输血前检查时发现抗体筛选阳性的送检标本样本,对其进行抗体鉴定试验确定抗体特异性均为抗-D。通过询问病史患者都是以往输用过Rh D阳性血液或不能明确所输用血液具体种类的个体。314例有过既往妊娠史的Rh阴性孕妇中产生抗-D46例,其中G2P0孕妇96例,产生抗-D个体4例,阳性率4.2%;G2P1孕妇41例,产生抗-D个体6例,阳性率14.6%;G3P0孕妇125例,产生抗-D个体10例,阳性率8.0%;G3P1孕妇38例,产生抗-D个体14例,阳性率36.8%;妊娠超过三次的孕妇38例,产生抗-D个体14例,阳性率36.8%。2、通过Dia Clon Rh-Subgroups+k卡测得对336例IAT阴性标本进行Rh表型鉴定。Ccee的个体为185例(55.06%),Ccee个体119例(35.42%),CCee个体14例(4.17%),cc Ee个体11例(3.27%),Cc Ee个体7例(2.08%)。3、通过Dia Clon Rh-Subgroups+k卡测得68例产生抗-D的Rh D阴性个体中,Rh表型为ccee的个体为56例(82.3%),Ccee个体5例(7.4%),CCee个体4例(5.9%),cc Ee个体2例(2.9%),Cc Ee个体1例(1.5%)。其中表型为ccee的个体产生抗-D的比例最高,达82.3%。4、通过人类红细胞RHD阴性鉴定基因检测试剂盒对336份样品进行检测,发现其中249例(74.1%)个体完全缺失RHD基因,l68例(20.2%)个体携带RHD1227A等位基因,19例(5.6%)携带RHD-CE(2-9)-D融合基因。5、68例RHD1227A个体Rh表型为ccee的个体为0例,Ccee个体56例(82.4%),CCee个体8例(14.2%),cc Ee个体1例(1.8%),Cc Ee个体3例(5.36%)。6、抗体鉴定确定产生Ig G抗-D的个体68例,其中63例(92.6%)完全缺失RHD基因,5例(7.4%)携带RHD-CE(2-9)-D融合基因,携带RHD1227A等位基因未检出产生抗-D的个体。产生抗体的基因型均为完全缺失RHD基因和RHD-CE(2-9)-D融合基因个体,DEL RHD1227A个体没有产生抗-D。结论血清学确认Rh D阴性个体的RHD基因型具有多态性,北京地区主要以RHD基因缺失为主,其次为RHD-CE(2-9)-D型和DEL RHD1227A。抗-D的产生与Rh阴性个体受到D抗原刺激的数量有关,妊娠和生育的次数与抗D的产生呈正相关。完全缺失RHD基因和RHD-CE(2-9)-D融合基因个体被D抗原免疫时有同种免疫风险,作为受血者应输用阴性血。Del个体与C抗原有高度相关性,在中国汉族人群中最常见的DEL型RHD1227A个体没有产生抗-D,或许可以输用阳性血。
[Abstract]:Objective to identify the RHD genotype of Rh D negative individuals by serological test, and to study the relationship between Rh D negative individuals with different genetic backgrounds and anti -D homoimmunization. The strategy of blood donation and blood transfusion for various Rh D variant individuals was focused on. Method 1, from March 2011 to 2013, 6 The Rh negative individuals in the monthly hospital were excluded by the history of blood transfusion and the history of pregnancy. By using saline test tube and indirect anti human ball test (IAT), the individuals who were positive in the serological test were excluded. The abortion history and birth history of pregnant women were classified, and the effects of abortion history and birth history against -D were studied, and.2 was studied. The antibody was screened by the microcolumn gel method and the antibody specificity was identified through the spectral cells. The Rh phenotypes of these samples were detected, including D, C, C, E and e antigen.3. Sequence specific primers PCR (PCR-SSP) and DNA sequence analysis techniques were used to analyze the RHD gene of the samples. Human erythrocyte RHD negative identification gene detection kit could be detected. The genotypes included Rh D positive, Rh D negative, Rh D-CE (2-9) -D, DVa (Hus), DVI III type, weak D15 type and the result of 1. In this study, there were 336 cases of immunological history negative individuals. The test determined that the specificity of the antibody was anti -D. by inquiring the patient history of the patients who had previously been transfused with Rh D positive blood or the specific type of blood of the individuals who were unable to specify the specific type of blood in the past pregnancy history of Rh negative pregnant women. Among them, 96 cases of G2P0 pregnant women, 4 cases of anti -D individuals, the positive rate of 4.2%, 41 cases of G2P1 pregnant women, were produced. There were 6 cases of anti -D, positive rate of 14.6%, 125 cases of G3P0 pregnant women, 10 cases of anti -D individual, 8% positive rate, 38 cases of pregnant women, 14 cases of anti -D individuals, 36.8% of the positive rate, 38 cases of pregnant women over three times, 14 cases of anti -D individuals, positive rate 36.8%.2, and Rh phenotypic identification of IAT negative specimens of 336 cases were tested by Dia Clon Rh-Subgroups+k card. Rh phenotypes were identified for IAT negative specimens of 336 cases by Dia Clon Rh-Subgroups+k card There were 185 individuals (55.06%), 119 Ccee individuals (35.42%), 14 CCee individuals (4.17%), 11 CC Ee individuals (3.27%), 7 Cc Ee individuals (2.08%).3, and 68 cases of -D Rh (82.3%) by Dia Clon Rh-Subgroups+k card. 2.9%) 1 cases (1.5%) of Cc Ee individuals. Among them, the proportion of individuals with CCEE was highest, reaching 82.3%.4. 336 samples were detected by human erythrocyte RHD negative identification gene detection kit, and 249 cases (74.1%) were completely missing RHD gene, L68 case (20.2%) carrying RHD1227A allele, 19 (5.6%) carrying RHD-. The Rh phenotype of CE (2-9) -D fusion gene was 0, 56 of Ccee individuals (82.4%), 8 in CCee (14.2%), 1 in CC Ee individual (1.8%), 3 in Cc Ee (5.36%). RHD1227A alleles were not detected to produce anti -D individuals. All the genotypes produced by the antibody were completely missing RHD gene and RHD-CE (2-9) -D fusion gene, and DEL RHD1227A individuals did not produce a -D. conclusion that serology confirmed Rh D negative individuals. The production of HD-CE (2-9) -D and DEL RHD1227A. resistance to -D was related to the number of Rh negative individuals stimulated by D antigen. The number of pregnancy and fertility was positively related to the production of anti D. The total deletion of RHD and RHD-CE (2-9) -D fusion genes were immune to the same immune risk when immunized with D antigen. Highly correlated, in Chinese Han population, the most common type DEL RHD1227A individuals did not produce anti -D and may lose positive blood.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6
本文编号：2145265
[Abstract]:Objective to identify the RHD genotype of Rh D negative individuals by serological test, and to study the relationship between Rh D negative individuals with different genetic backgrounds and anti -D homoimmunization. The strategy of blood donation and blood transfusion for various Rh D variant individuals was focused on. Method 1, from March 2011 to 2013, 6 The Rh negative individuals in the monthly hospital were excluded by the history of blood transfusion and the history of pregnancy. By using saline test tube and indirect anti human ball test (IAT), the individuals who were positive in the serological test were excluded. The abortion history and birth history of pregnant women were classified, and the effects of abortion history and birth history against -D were studied, and.2 was studied. The antibody was screened by the microcolumn gel method and the antibody specificity was identified through the spectral cells. The Rh phenotypes of these samples were detected, including D, C, C, E and e antigen.3. Sequence specific primers PCR (PCR-SSP) and DNA sequence analysis techniques were used to analyze the RHD gene of the samples. Human erythrocyte RHD negative identification gene detection kit could be detected. The genotypes included Rh D positive, Rh D negative, Rh D-CE (2-9) -D, DVa (Hus), DVI III type, weak D15 type and the result of 1. In this study, there were 336 cases of immunological history negative individuals. The test determined that the specificity of the antibody was anti -D. by inquiring the patient history of the patients who had previously been transfused with Rh D positive blood or the specific type of blood of the individuals who were unable to specify the specific type of blood in the past pregnancy history of Rh negative pregnant women. Among them, 96 cases of G2P0 pregnant women, 4 cases of anti -D individuals, the positive rate of 4.2%, 41 cases of G2P1 pregnant women, were produced. There were 6 cases of anti -D, positive rate of 14.6%, 125 cases of G3P0 pregnant women, 10 cases of anti -D individual, 8% positive rate, 38 cases of pregnant women, 14 cases of anti -D individuals, 36.8% of the positive rate, 38 cases of pregnant women over three times, 14 cases of anti -D individuals, positive rate 36.8%.2, and Rh phenotypic identification of IAT negative specimens of 336 cases were tested by Dia Clon Rh-Subgroups+k card. Rh phenotypes were identified for IAT negative specimens of 336 cases by Dia Clon Rh-Subgroups+k card There were 185 individuals (55.06%), 119 Ccee individuals (35.42%), 14 CCee individuals (4.17%), 11 CC Ee individuals (3.27%), 7 Cc Ee individuals (2.08%).3, and 68 cases of -D Rh (82.3%) by Dia Clon Rh-Subgroups+k card. 2.9%) 1 cases (1.5%) of Cc Ee individuals. Among them, the proportion of individuals with CCEE was highest, reaching 82.3%.4. 336 samples were detected by human erythrocyte RHD negative identification gene detection kit, and 249 cases (74.1%) were completely missing RHD gene, L68 case (20.2%) carrying RHD1227A allele, 19 (5.6%) carrying RHD-. The Rh phenotype of CE (2-9) -D fusion gene was 0, 56 of Ccee individuals (82.4%), 8 in CCee (14.2%), 1 in CC Ee individual (1.8%), 3 in Cc Ee (5.36%). RHD1227A alleles were not detected to produce anti -D individuals. All the genotypes produced by the antibody were completely missing RHD gene and RHD-CE (2-9) -D fusion gene, and DEL RHD1227A individuals did not produce a -D. conclusion that serology confirmed Rh D negative individuals. The production of HD-CE (2-9) -D and DEL RHD1227A. resistance to -D was related to the number of Rh negative individuals stimulated by D antigen. The number of pregnancy and fertility was positively related to the production of anti D. The total deletion of RHD and RHD-CE (2-9) -D fusion genes were immune to the same immune risk when immunized with D antigen. Highly correlated, in Chinese Han population, the most common type DEL RHD1227A individuals did not produce anti -D and may lose positive blood.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6
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