[Application of denaturing high performance liquid chromatography for the detection of maternal DNA contamination during prenatal diagnosis]
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Microsatellite DNA Genotyping and Flow Cytometry Ploidy Analyses of Formalin-fixed Paraffin-embedded Hydatidiform Molar Tissues
Published on: October 20, 2019
Lijie Gao1, Yanming Long, Rong Zhang
1Huizhou Second Maternal and Child Health Hospital, Huizhou, Guangdong 516001, P.R.China. guoping_he@163.com.
Objective:
To establish a high-quality method for detecting short tandem repeats(STR) using denaturing high performance liquid chromatography(DHPLC) in order to exclude maternal contamination and improve the accuracy of prenatal diagnosis.
Methods:
Two families were recruited. DNA was extracted from blood samples from the parents as well as amniotic fluid. Sixteen STR sites were amplified and analyzed based on the range of allele length reported by a STR database. Maternal DNA was mixed with DNA derived from amniotic fluid samples with the ratio 1:1, 1:4, 1:9, 1:19 and 1:99. vWA STR site was detected with DHPLC to confirm the sensitivity of detection.
Results:
Sixteen STR sites were analyzed by DHPLC, for which at least 10 were found to be different between the mothers and fetuses. The detection rate, with maternal contamination excluded, was 66.7%. And the sensitivity of detection was 1-10%.
Conclusion:
Maternal contamination of amniotic fluid can be rapidly excluded with accuracy with DHPLC, which features a high sensitivity and good quality control, and can meet the European standards and provide a reliable quality control platform for prenatal diagnosis.
