Whole Genomic Amplification for Detecting Single-Gene Variants Causing Inherited Disease

Area of Science:

• Reproductive Medicine
• Genetics
• Embryology

Background:

• Whole genome amplification (WGA) is crucial for genetic analysis using limited DNA samples.
• Preimplantation Genetic Testing (PGT) aids in assisted reproduction by screening embryos for genetic abnormalities.
• PGT includes testing for aneuploidy (PGT-A) and specific monogenic diseases (PGT-M).

Purpose of the Study:

• To detail the WGA process for clinical applications in preimplantation genetic diagnosis.
• To demonstrate the utility of WGA in PGT-M for Mendelian diseases.
• To showcase WGA's application in embryo sex diagnosis via Y chromosome detection.

Main Methods:

• Whole genome amplification (WGA) protocol optimization.
• Polymerase chain reaction (PCR) fragment preparation for Sanger sequencing.
• Y chromosome detection assays for sex determination.

Main Results:

• Successful WGA enabling detection of Mendelian diseases in embryos.
• Identification of specific genetic variants for healthy embryo selection in IVF.
• Accurate detection of the Y chromosome for sex diagnosis in embryos.

Conclusions:

• WGA is a valuable tool for PGT-M, facilitating the conception of offspring free from specific monogenic diseases.
• WGA combined with PGT-A and PGT-M enhances the success of in vitro fertilization (IVF) treatments.
• The WGA method supports comprehensive genetic screening of embryos, including disease diagnosis and sex determination.