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Enrichment, Identification and Isolation of Fetal Nucleated Red Blood Cells from Maternal Blood

Author: ZhouYun
Tutor: LiuYuSheng
School: Anhui Medical University,
Course: Obstetrics and Gynaecology
Keywords: Fetal nucleated red blood Transferrin receptor Leukocyte common antigen Flow cytometry Prenatal diagnosis Hemoglobin ε chain
CLC: R714.5
Type: Master's thesis
Year: 2008
Downloads: 58
Quote: 0
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Abstract


The purpose of the use of anti- transferrin receptor (CD71) antibody and anti- double-labeled leukocyte common antigen (CD45) antibody assay gestational age the proportion of nucleated red blood cells in the peripheral blood of pregnant women to choose the best weeks of gestation fetal nucleated red blood cells enriched . Method using anti- CD71 antibody and anti-CD45 antibody double labeling using flow cytometry (flow cytometry, FCM) to detect the proportion of nucleated red blood cells in the peripheral blood of 81 cases of pregnant women in different gestational weeks , compared with the nonpregnant group and cord blood group . CD71 / CD45dim cells in peripheral blood ratio of 7 to 34 weeks of pregnancy results pregnant (0.107 ± 0.063) × 10-2, compared to non-pregnant group and cord blood group differences were statistically significant ; CD71 / CD45dim cells in peripheral blood in the proportion of pregnant 17 ~ 18 Zhou Dafeng value ( 0.144 ± 0.096 ) × 10-2 . The conclusion is different gestational age of the maternal blood nucleated red blood cells in different proportions , the highest proportion of 17 to 18 weeks gestation . The best time of fetal nucleated red blood cells in the maternal circulation for prenatal diagnosis of selectable in 17 to 18 weeks of pregnancy . Purpose enriched in maternal blood fetal nucleated red blood cells ( fetal nucleated red blood cells , FNRBCs ) using flow cytometry identify maternal blood ε- hemoglobin chain (epsilon-haemoglobin-chain, Hbε) positive nucleated red blood cells (NRBCs) in order to non - invasive prenatal diagnosis , and finding the best detection gestational age . The use of density gradient method to extract nuclear cell layer enriched in 10 to 25 weeks of gestation in maternal blood FNRBCs by flow cytometry using CD71 antibody staining , then the use of Hbε antibody staining the identification of the fluorescence microscope Hbε positive NRBCs . The results identified in the 14 cases , 10 to 15 weeks' gestation , maternal blood Hbε the positive NRBCs13 cases , the positive rate was 92.86% , the positive samples per 10ml maternal blood in an average of 6.92 ± 2.75 Hbε positive NRBCs . Identified 12 cases of 16 to 25 weeks of gestation , maternal blood out Hbε positive NRBCs 9 cases , the positive rate was 75.00% , the positive samples per 10ml maternal blood in an average of 2.67 ± 1.32 Hbε positive NRBCs . Conclusion The use of flow cytometry enriched maternal blood FNRBCs using Hbε antibodies can the identified Hbε positive NRBCs the antibody recognition applications Hbε FNRBCs best period is selectable in 10 to 15 weeks' gestation .

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CLC: > Medicine, health > Obstetrics and Gynaecology > Obstetrics > Fetus
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