Fetal gender determination through Y-STR analysis of maternal plasma during the third trimester of pregnancy

Hanaa M.H Aal-Hamdan,Ahmed M Refaat,Saranya R Babu,Abdul Rauf Choudhry

doi:10.1016/j.ejmhg.2014.08.005

Hanaa M.H Aal-Hamdan, Ahmed M Refaat + Show 2 more

Open Access

https://doi.org/10.1016/j.ejmhg.2014.08.005

Copy DOI

Abstract

BackgroundThe passage of nucleated cells between mother and fetus is well recognized (Lo et al., 1989, 1996). As well as, cell-free fetal DNA in maternal plasma or serum is at present widely investigated as a source of fetal genetic material (Stanghellini et al., 2006) [18]. There has been much recent interest in the use of DNA derived from plasma or serum (Boland, 1996). This DNA can be utilized for molecular diagnosis as well as prenatal sex discernment. ObjectiveTo establish an easy, reliable, and completely safe method for fetal gender determination alternative to conventional exhausting current techniques applied in gynecologic hospitals and clinics, besides its further applications in forensic casework. MethodsEDTA-Blood samples were taken from 30 pregnant women all in the third trimester of pregnancy, then plasma was separated from each sample, from which DNA was isolated using a QIAamp DNA Mini Kit, with special modifications done in the extracting procedure to concentrate and obtain minute quantities of fetal DNA, together with maternal DNA, from maternal plasma. In addition, bloodstain samples were taken from the husbands of women who were pregnant with male fetuses from which DNA was isolated using a QIAamp DNA Micro Kit for comparison. DNA quantification was done using a Real-time PCR utilizing Quantifiler Duo Kit. PCR was done using an AmpFlSTR Y-Filer Kit, then amplified products were typed using a 3130 Genetic Analyzer. ResultsFull and partial Y-STR profiles (6–17 STR loci) were obtained from all plasma samples taken from pregnant women with male fetuses, while negative Y-STR profiles (no single STR locus) were obtained from all plasma samples taken from pregnant women with female fetuses. ConclusionIt is recommended to use Y-STR profiling as an alternative technique for fetal gender determination during the third trimester of pregnancy, in addition to its significance in forensic casework.

Highlights

Prenatal sex discernment is the prenatal testing for discerning the sex of a fetus before birth
EDTA-Blood samples were taken from 30 pregnant women all in the third trimester of pregnancy, plasma was separated from each sample, from which DNA was isolated using a QIAamp DNA Mini Kit, with special modifications done in the extracting procedure to concentrate and obtain minute quantities of fetal DNA, together with maternal DNA, from maternal plasma
Concentration concentration maternal plasma samples from the 30 women included in the study during pregnancy, and in the second time it was done for the DNA extracted from the 12 bloodstain samples from the husbands of the women who were pregnant with male fetuses after delivery

Summary

Introduction

Prenatal sex discernment is the prenatal testing for discerning the sex of a fetus before birth. Chorionic villus sampling (CVS) and amniocentesis are two rather invasive testing procedures. The difficulty of these tests and the risk of damage to the fetus, potentially result in miscarriage or congenital abnormalities. Either transvaginally or transabdominally, can check for the sagittal sign as a marker of fetal sex. It gives a result in 90% of cases [1]. There has been much recent interest in the use of DNA derived from plasma or serum (Boland, 1996) This DNA can be utilized for molecular diagnosis as well as prenatal sex discernment

Methods

Results

Conclusion