Analysis of fetal sex and RhD gene in fetal cells DNA from maternal blood by polymerase chain reaction
Objective: The Purpose of this study was to determine fetal nucleated red blood cells (NRBCs) among enriched maternal peripheral blood and to evaluate the effectiveness of enriching NRBCs using nested-PCR methods. Material and methods: We obtained approximately 30ml blood from 53 pregnant women. The mononuclear cells were isolated with ficole. To remove unwanted maternal white blood cells, magneticbeads coated with antibodies against CD45 and CD14 were used as negative selection. Positive selection methods based on using antibody against CD71 and Dynal magnetic were employed to enrich fetal nucleated erythrocytes. DNA was extracted from fetal cellsby a standard phenol-chloroform procedure. Nested PCR specific for a region of the amelogenine gene was used for Sex (X and Y) determination as well as for RhD gene. Results: The most interesting data obtained by isolation of the nucleated fetal erythrocyes. At the DNA level in order to detect fetal sex and RhD PCR was performed to identify fetal sex and RhD in maternal blood circulation in 27 (51%) out of 53 cases. Conclusion: These results suggest that identification of fetal cells sex and RhD in the maternal blood circulation is possible.