| Objective:To detect free fetal DNA from maternal peripheral blood , explore the possibility of prenatal gene diagnosis for fetal Rh blood group with free fetal DNA and search a new method of non-invasive prenatal gene diagnosis . Methods:(1)A single-copy human DYS14 gene of Y-chromosome of fetal DNA sequence was amplified from 170 pregnant plasma or serum samples (between12?0 weeks) by nested polymerase chain reaction (PCR);(2)The RHD gene was amplified from 13 RhD-negative pregnant plasma ( between 18~20 week ) by nested PCR .The maternal plasma and serum samples of 183 pregnant women were used directly for nested PCR. Results:(1)The DYS14 gene electrophoresis bands were identified in 108 of 140 male-bearing pregnant women. 17 of 75 maternal plasma samples had no the electrophoresis bands, the diagnostic accordance rate was 77.3%(58/75).15 of 65 maternal serum samples had no electrophoresis bands. The diagnostic accordance rate was 76. 9%(50/65), 2 of 30 female-bearing pregnant blood samples had the electrophoresis bands. The final accuracy of 80.0% was attained in all cases; (2) The RH gene electrophoresis bands were identified in 7 of 10 fetal Rh blood group positive pregnant women, The diagnostic accordance rate was 70%, none of 3 fetal Rh blood group negative pregnant women had the electrophoresis bands. The final accuracy of 77.0% was attained in all cases. Conclusions: 0Fetal free DNA is present in maternal plasma and serum. The detecting rate of fetal DNA in maternal peripheral blood increased with gestational ages; (2) Fetal free DNA in maternal peripheral blood can be used for prenatal diagnosis of Rh blood group incompetibility;(3)The nested PCR that detects fetal free DNA from peripheral blood possesses the advantages of sensitivity, sepcificity and detecting rate. It can be used for clinic as a non-invasive prenatal gene diagnostic method.Postgreduate Chang jing (Genetics)Directed by Wang peilin Wang xiuhai...
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