A Study On Their Rhd Genotypes Of Rh Negative Pregnant Women And The Relationship Between Genotypes And Anti-d Antibody

Haemolytic Disease of the Newborn (HDN) mainly due to maternal and child's blood group incompatibility, Rh HDN mainly occur in RhD-negative the mothers who have a RhD-positive fetus,that HDN caused by RhD blood group incompatibility. So during RhD-negative women are pregnancy, the fetal RhD blood group is detected. It is great significance for the prevention, diagnosis Rh HDN. Now before birth fetal RhD gene is detected with PCR method from maternal blood or amniotic fluid and then infer RhD blood group. This requires the high degree of consistency between RhD genotype and phenotype or the fixed link can be established each other. However, because RhD-negative Chinese show polymorphism of D exon, the methods that are builted on the basis of the Caucasian RhD qualitative gene can not be applied in these cases. So to find the relationship RhD between genotype and phenotype appears to be extremely important to in China through implemente that Rh phenotype infer from Rh gene. At the same time, in clinical blood transfusion, who RhD-negative receive serological RhD-positive blood produce the same kind of immunity only 50-75%, and RhD-negative pregnant women befell HDN only 5% -7%, it explain that not all of the RhD- negative would have the same-kind immunity. On the one hand, a result of this study was to explore the relationship of RHD genotype and phenotype so that the fetal genotype can be to some extent inferred from its phenotype prenatal diagnosis; on the other hand, to analyze the relationship between genotypes and anti-D antibodies in the RHD - negative pregnant women ,it provide the theoretical guidance to clinical prediction and prevention of hemolytic disease of the newborn.Part oneObjective To investigate RhD-negative pregnant women' with in Zhongyuan region genotypes and phenotypes and to find out the relationship between genotypes and phenotypes. In order to provide a theoretical basis that RhD phenotype inferred from the detection of fetal RHD gene and it is very meaningful for the clinical diagnosis and the prevention of RhD HDN.Methods 70 cases of RhD-negative pregnant women were selected with the hemagglutination test .Every specimen of various RhCcEe serotype was tested with the appropriate standard serums and was tested RhD-classification with absorbing-emission test and Indirect Antiglobulin Test (ITA) ;The polymerase chain reaction-sequence specific oligonucleotide primer (PCR-SSP) was used to amplify the RHD,RHC/E gene exon in 70 samples . Comparatively to analyze the relationship between genotypes and phenotypes.Resultsâ‘ Phenotype frequence of Rh c,e,C,E in 70 pregnant women with RhD-negative were respectively 25.71 % ,74.29 % ,7.14 % and 92.86 % with the hemagglutination test; 54 cases in 70 cases were true-negative, no weak was positive, 16 cases were del with absorbing-emission test and Indirect Antiglobulin Testâ‘¡RHC/E genotype were exactly the same to phenotype;RHD genes of RhD-negative individuals with C antigens have polymorphism. In 70 cases of RhD serology negative individuals with RhC/E typing for CC or Cc were 28,of which 8 cases carried intact exons ,4 cases carried partial deletion exons,and 16 was total deletion; The individuals without C antigen (phenotype of ccee ,ccEe or ccEE)was total deletion;â‘¢15 cases of 16 Del were no deletion of the RHD gene exons. only 1 case carried partial deletion exons.Conclusion .â‘ RHC/E genotypes were exactly the same to its phenotypes ,so their genotypes can be determined in accordance with their phenotypes. Therefore, fetal RhC / E phenotype can be inferred from its C / E genotype during prenatal diagnosis;â‘¡gene structures of the individuals with C gene have polymorphism,which are accorded with the Oriental character:no deletion,partial deletion and total deletion. Gene structures of the individuals without C gene are total deletion .Therefore, if the prenatal diagnosis of fetal RHD gene is no deletion and it can not determine its phenotype is RhD-positiveâ‘¢RHD exon of Del is mainly no deletion.Part twoObjective To check out those that anti-D antibody is positive to analyze the influence on antibody D of their abortion history, reproductive history .Simultaneity to analyze their genotype and phenotype in order to find out the rule the pregnant women with RhD-negative produce antibody-D. And to assess of the risk of antibodies from the genotype or phenotypes preliminarily.Methods Their anti-D antibody were filtered and their titer was determinationed with micro-column gel irregular antibody screening cards and pregnant women' titer determination of anti-D cards; for pregnant women at the same time statistical classifications of the history of abortion and reproductive history were gone on to research the influence on the production and titer of anti-D of the history of abortion, reproductive history. Antigen RhD was further classified with Indirect Antiglobulin Test and Absorption- emission Test ; C / E phenotypes were tested with the coagulation tests; genotypes of samples whith anti-D were positive were examined using PCR-SSP technology.Resultsâ‘ With micro-column gel irregular antibody screening and pregnant women' titer determination of anti-D cards in RhD (-) pregnant women whose anti-D total were positive were 35 cases, of which 6 cases titer were greater than or equal to 512, 3 cases were 256,8 cases were 128,6 cases of patients with 64,4 cases were 32,5 cases were 16,3 cases were 8. The positive cases were those who had abortions or childbirth; the more numbers of abortion or childbirth ,the greater chance of antibodies occur, the higher the price effect appear ; at the same time reproductive history are more likely to produce antibodies than the history of abortion ; the emergence of anti-D and anti-D titer is closely related to the history of abortion, reproductive history .Relative to the history of abortion, reproductive history was more relevant to the emergence;â‘¡35 cases were tested Absorption- emission Testwhich were all true-negative, no 1 for Del.â‘¢RhC / E phenotypes of 35 cases ofanti-D-positive pregnant women were main ccee with the coagulation tests ;â‘£No one of 35 cases was no deletion of RhD gene exons detected by PCR-SSP.Conclusionâ‘ The emergence of anti-D and anti-D titer is closely related to the history of abortion, reproductive history .Relative to the history of abortion, reproductive history was more relevant to the emergence of anti-D;â‘¡The positive cases of anti-D were mostly the category of total deletion of RhD gene exons and were serological ccee,. However, no one was no deletion and no one was Del.