Practicability of prenatal testing using lectin‐based enrichment of fetal erythroblasts. Issue 8 (2nd May 2016)

• Record Type: Journal Article
• Authors: Kanda, Eriko; Yura, Hirofumi; Kitagawa, Michihiro
• Is Part Of: Journal of obstetrics and gynaecology research. Volume 42:Issue 8(2016)
• Page Start: 918
• Page End: 926
• Publication Date: 2016-05-02
• Subjects: erythroblast -- fluorescence in situ hybridization -- lectin -- prenatal diagnosis -- sex‐determining region Y gene; Gynecology -- Periodicals; Obstetrics -- Periodicals; 618.1005
• DOI: 10.1111/jog.12982
• Languages: English
• ISSNs: 1341-8076

Abstract

Abstract: Aim: The aim of this study was to investigate the practicability and efficiency of lectin‐based isolation of fetal erythroblasts for clinical use in non‐invasive prenatal testing. Methods: Peripheral blood samples were collected from 39 pregnant women. Leukocytes were removed with an anti‐CD45 antibody after density gradient centrifugation. After blood cells were attached to slides by binding to a galactose‐specific lectin and galactose‐bound vinyl polymer, the slides were stained with May–Grünwald–Giemsa stain and cells were classified by automated image analysis based on their size and the nuclear area/cytoplasmic area ratio. In 14 samples from the women with male fetuses, fetal origin of the isolated erythroblasts was confirmed by detecting the Y chromosome using fluorescence in situ hybridization. In eight samples, single erythroblasts were collected by the laser capture microdissection technique for amplification of the sex‐determining region Y gene to confirm fetal origin. Results: Panning with an anti‐CD45 antibody achieved stable removal of leukocytes without aggregation. In all samples, erythroblasts were successfully identified by automated image analysis (18–6000/10 mL of blood). The number of slides required to examine 10 mL of blood ranged from one to six, which was reasonable for clinical use. The Y chromosome was detected in 7.5–43.6% of erythroblasts by fluorescence in situ hybridization, and the sex‐determining region Y gene was amplified in seven of eight samples. Conclusion: The combination of lectin‐based erythroblast isolation and automated image analysis is a practical and efficient method for isolating fetal erythroblasts as a source of fetal genomes.