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Published on: October 20, 2019
Preimplantation diagnosis for long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency
Yury Verlinsky1, Svetlana Rechitsky, Oleg Verlinsky
1836 West Wellington Avenue, Chicago, IL 60657, USA.
Insights
Preimplantation genetic diagnosis (PGD) successfully identified mutation-free oocytes for Long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency. This enabled the birth of a healthy child, avoiding termination of pregnancy.
Area of Science:
- Genetics
- Reproductive Medicine
- Metabolic Disorders
Background:
- Long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency is a fatal, inherited metabolic disorder.
- Preimplantation genetic diagnosis (PGD) offers a method to establish unaffected pregnancies, circumventing prenatal diagnosis and potential termination.
- In-vitro fertilization (IVF) techniques are crucial for implementing PGD.
Purpose of the Study:
- To develop and validate a method for pre-selecting mutation-free oocytes for LCHAD deficiency using PGD.
- To assess the efficacy of polar body testing in identifying unaffected oocytes.
- To establish an unaffected pregnancy through embryo transfer derived from pre-selected oocytes.
Main Methods:
- Micromanipulation techniques were used to remove and test the first and second polar bodies from oocytes.
- Hemi-nested polymerase chain reaction (PCR) was employed for mutation detection to prevent misdiagnosis.
- Outer primers were specifically designed to avoid pseudogene amplification, ensuring accurate results.
Main Results:
- Out of 12 tested oocytes, 4 were predicted to be unaffected based on polar body analysis.
- Embryos derived from these mutation-free oocytes were transferred.
- A singleton clinical pregnancy was achieved, resulting in the birth of a healthy child.
Conclusions:
- PGD utilizing polar body testing is a viable strategy for preventing the transmission of LCHAD deficiency.
- This approach allows for the establishment of unaffected pregnancies, offering a significant reproductive option.
- The hemi-nested PCR method with specific primer design enhances diagnostic accuracy in PGD for genetic disorders.
Abstract:
Long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency is a fatal autosomal recessive metabolic disorder, presenting during infancy. Preimplantation genetic diagnosis (PGD) provides an option for establishing an unaffected pregnancy, avoiding the risk for termination of pregnancy following prenatal diagnosis. The method for pre-selection of mutation-free oocytes for LCHAD deficiency was developed by testing the first and second polar body removed from oocytes by micromanipulation techniques in the framework of in-vitro fertilization. To avoid misdiagnosis, testing was done using hemi-nested polymerase chain reaction (PCR), with outer primers designed to lie outside the pseudogene, eliminating false priming. Four of 12 tested oocytes were predicted to be unaffected, based on the heterozygous first and mutant second polar body. The embryos resulting from these mutation-free oocytes were replaced, yielding a singleton clinical pregnancy and birth of a healthy child following confirmation by prenatal diagnosis.
