Detection of fetal nucleated erythrocytes and fetal DNA from peripheral blood of pregnant women

Abstract

In order to search for a more reliable method of sorting fetal nucleated red blood cells (NRBCs) and DNA from maternal peripheral blood and to identify origin of NRBCs and DNA, NRBCs were isolated from peripheral blood of 88 pregnant women by density gradient centrifugation and fluorescence activated cell sorter (FACS) respectively. Nested polymerase chain reaction was used to detect normal male SRY gene from blood plasma DNA of 65 pregnant women. The results revealed that fetal NRBCs were found in 14 of 27 maternal samples by density gradient centrifugation. The number of cells was from 1 to 10. Using FACS, CD71⁺ cells were identified among all 61 samples. The frequency was (0.35±0.25)×10⁻²; The detectable rate of the SRY gene of blood plasma DNA from 46 women carrying male fetuses was 65.22% (30/46). Non-detectable rate for 19 women carrying female fetuses was 94.74% (18/19). It was concluded that the methods of sorting fetal NRBSs and DNA have already made great progress. The methods for fetal NRBCs and plasma DNA from maternal peripheral blood to diagnose genetic diseases seem to be the best methods of noninvasive prenatal diagnosis.

Keywords

nucleated red blood cells / fetal DNA / prenatal diagnosis

References

[1] MachtelS S, SammonsD, TwittyG, et al.. Charge flow separation: quantification of nucleated red blood cells in maternal blood during pregnancy. Prenat Diagn, 1998, 18(5): 455-455

[2] OosterwijkJ C, KnepfleC F, MeskerW E, et al.. Strategies for rare-event detection: an approach for automated fetal cell detection in maternal blood. Am J Hum Genet, 1998, 63(6): 1783-1783

[3] ChenH, FriffinD K, JesticeK, et al.. Evaluating the culture of fetal erythroblasts from maternal blood for non-invasive prenatal diagnosis. Prenat Diagn, 1998, 18(9): 883-883

[4] BianchiV W, KlingerK W, VadnaisT J, et al.. Development of a model system to compare all separation methods for the isolation of fetal cells from maternal blood. Prenat Diagn, 1996, 16(4): 289-289

[5] TakabayashiH, KuwabaraS, UkitaT, et al.. Development of non-invasive fetal DNA diagnosis from maternal blood. Prenat Diagn, 1995, 15(1): 74-74

[6] ThilaganathanB, Meher-HomjiN J, NicolaidesK H. Blood transferrin receptor expression in chromosomally abnormal fetuses. Prenat Diagn, 1995, 15(3): 282-282

[7] BianchiD W, WilliamsJ M, SullivanL M, et al.. PCR quantitation of fetal cells in maternal blood in normal and aneuploid pregnancies. Am J Hum Genet, 1997, 61(4): 822-822

[8] CheungM C, JamesD G, YuetW K, et al.. Prenatal diagnosis of sickle cell anaemia and thalassaemia by analysis of fetal cells in maternal blood. Nature Genet, 1996, 14(3): 264-264

[9] SekizawaA, WatanabeA, KimuraT, et al.. Prenatal diagnosis of fetal RhD blood type using a single fetal nucleated erythrocyte from maternal blood. Obstet Gynecol, 1996, 87(4): 501-501