Restriction enzyme-mediated enhanced detection of circulating cell-free fetal DNA in maternal plasma

摘要: A universal method confirming the presence of circulating cell-free fetal (ccff) DNA in maternal plasma is important field noninvasive prenatal diagnostics. Restriction endonuclease digestion one allele a single-nucleotide polymorphism (SNP) was used to allow detection paternal alleles DNA. Multiplexed genotyping 92 panethnic high-frequency SNPs predicted >0.99 probability detecting at least four informative loci per sample. Child-maternal paired samples were confirm 2% child's heterozygous background homozygous for digestible allele. By restriction PCR cocktail before thermal cycling, 10 genomic copies SNP detectable 990 alleles. comparison 154 pregnant and nonpregnant female demonstrated enhanced nondigestible plasma. Receiver operating characteristic curve analysis showed an optimal threshold confirmation ccff 98% sensitivity 96% specificity 95% confidence level. Our study demonstrates ability this technique from