Identification of a Novel Single Nucleotide Polymorphism of HADHA Gene at a Referred Primer-binding Site During Pre-diagnostic Tests for Preimplantation Genetic Diagnosis.
10.3346/jkms.2006.21.5.794
- Author:
Hyoung Song LEE
1
;
Hye Won CHOI
;
Chun Kyu LIM
;
Mi Kyoung KOONG
;
Inn Soo KANG
;
Han Wook YOO
;
Jin Ho CHOI
;
Jin Hyun JUN
Author Information
1. Laboratory of Reproductive Biology and Infertility, Cheil General Hospital and Women's Healthcare Center, Sungkyunkwan University School of Medicine, Seoul, Korea. junjh55@hotmail.com
- Publication Type:Original Article
- Keywords:
Preimplantation Diagnosis;
long-chain 3-hydroxyacyl CoA dehydrogenase;
LCHAD Deficiency;
peroxysomal bifunctional enzyme;
HADHA Gene;
Allele Drop-out;
Polymorphism, Single Nucleotide
- MeSH:
*Preimplantation Diagnosis;
*Polymorphism, Single Nucleotide;
Polymerase Chain Reaction;
Mutation;
Multienzyme Complexes/*genetics;
Male;
Humans;
Female;
Binding Sites;
Adult;
3-Hydroxyacyl CoA Dehydrogenases/deficiency
- From:Journal of Korean Medical Science
2006;21(5):794-799
- CountryRepublic of Korea
- Language:English
-
Abstract:
The pre-diagnostic test for preimplantation genetic diagnosis (PGD) of long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency was performed by polymerase chain reaction (PCR) and direct sequencing for hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A hydratase (HADHA) gene. We obtained unexpected genotyping results of HADHA gene by allele drop-out in the analysis of patients' genomic DNA samples with a referred PCR primer set. Upon further analysis with a re-designed primer set, we found a novel single nucleotide polymorphism (SNP) at the referred primer-binding site in the normal allele of HADHA gene (NT_022184, 5233296 a>t). We found that the frequency of this novel SNP was 0.064 in Korean population. Pre-diagnostic test using single lymphocytes and clinical PGD were successfully performed with the re-designed primer set. Nineteen embryos (95.0%) among 20 were successfully diagnosed to 5 homozygous mutated, 8 heterozygous carrier and 6 wild type. Among 6 normal embryos, well developed and selected 4 embryos were transferred into the mother's uterus, but a pregnancy was not achieved. We proposed that an unknown SNP at primer-binding sites would be a major cause of allele drop-out in the PGD for single gene dis-order.
