Performance of PCR method on Amelogenin gene in maternal blood for sex determination of embryos

Shirali, S.; Khodadi, E.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p16
Academic Journal
Introduction: Sex determination can not only be used for forensics investigations but can also be useful in prenatal diagnosis of fetus x-linked diseases and diagnosis of fetal abnormalities. Recently, the molecular techniques has special place and are highly important to determine gender. PCR methods using sequences that are both in X and Y chromosomes, such as ZFY and ZFX, Steroid Sulfatase (STS), Amelogenin gene, or only are on the Y chromosome like: SRY, DYS14, DYS1. Techniques used for sampling of the fetus are divided into two groups including invasive methods such as, chorionic villus sampling (CVS), Amniocentesis Precutaneous umblical blood sampling (PUBS), Fetal tissue sampling (FTS) methods and noninvasive methods that includes the detection of cell-free fetal DNA in serum or plasma. Materials and Methods: Maternal blood samples were collected and DNA was extracted from blood samples using modified phenol-chloroform extraction procedure. The primers of the Amelogenin gene were designed to yield products that would be readily distinguishable by their sizes upon agarose gel electrophoresis. Results: All of amplified samples which had been shown a single 467-bp fragment were interpreted as female and the others involved both the 467 and 341 bp amplification products were interpreted as male. This method could detect accurately the sex of male and females embryos by 90%. Conclusion: This assay provides a rapid and sensitive method for gender determination. The success of the procedure can be monitored by the presence of the X-chromosome band. The method contains an internal control which detects PCR failure. Two important merits of this research are its feasibility and reliability. All of these processes can be carried out in a regular laboratory or at an outside workstation and results are available within ten hours. By using PCR method on Amelogenin gene favorable results for sex determination of the fetus in the first trimester of pregnancy were obtained.


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