Application of short tandem repeat based linkage analysis in preimplantation genetic testing for Pompe disease

Abstract

Pompe disease is an autosomal recessive glycogen storage disorder caused by genetic alterations in the GAA gene, leading to a deficiency of the enzyme acid alpha-glucosidase. Preimplantation genetic testing for monogenic disorders (PGT-M) is an effective method for the prevention of Pompe disease. Six short tandem repeats (STR) markers (D17S802, D17S1806, D17S784, D17S78.0, D17S78.57, and D17S928) flanking the GAA gene were amplified using Multiplex polymerase chain reaction (Multiplex-PCR) for linkage analysis in 17 family members and 23 biopsied embryo samples from five families with a history of Pompe disease. Linkage analysis results were compared with Sanger sequencing. Linkage analysis and Sanger sequencing results were consistent in 22 out of 23 embryo samples (95.65%), except for one embryo (4.35%) that was misdiagnosed directly due to allelic dropout. Linkage analysis using a panel of 6 STRs near the GAA gene amplified by Multiplex-PCR has been successfully applied in PGT-M for Pompe disease. This technique is highly reliable, helps reduce the risk of misdiagnosis due to allelic dropout, and is compatible with current clinical practice in Vietnam.