Complex preimplantation genetic tests for Robertsonian translocation, HLA, and X-linked hyper IgM syndrome caused by a novel mutation of CD40LG gene

J Assist Reprod Genet. 2020 Aug;37(8):2025-2031. doi: 10.1007/s10815-020-01846-y. Epub 2020 Jun 5.


Purpose: To perform complex preimplantation genetic tests (PGT) for aneuploidy screening, Robertsonian translocation, HLA-matching, and X-linked hyper IgM syndrome (XHIGM) caused by a novel mutation c.156 G>T of CD40LG gene.

Methods: Reverse transcription PCR (RT-PCR) and Sanger sequencing were carried out to confirm the causative variant of CD40LG gene in the proband and parents. Day 5 and D6 blastocysts, obtained by in vitro fertilization (IVF) with intracytoplasmic sperm injection, underwent trophectoderm (TE) biopsy and whole genomic amplification (WGA) and next generation sequencing (NGS)-based PGT to detect the presence of a maternal CD40LG mutation, aneuploidy, Robertsonian translocation carrier, and human leukocyte antigen (HLA) haplotype.

Results: Sanger sequencing data of the genomic DNA showed that the proband has a hemizygous variant of c. 156 G>T in the CD40LG gene, while his mother has a heterozygous variant at the same position. Complementary DNA (cDNA) of CD40LG amplification and sequencing displayed that no cDNA of CD40LG was found in proband, while only wild-type cDNA of CD40LG was amplified in the mother. PGT results showed that only one of the six tested embryos is free of the variant c.156 G>T and aneuploidy and having the consistent HLA type as the proband. Meanwhile, the embryo is a Robertsonian translocation carrier. The embryo was transplanted into the mother's uterus. Amniotic fluid testing results are consistent with that of PGT. A healthy baby girl was delivered, and the peripheral blood testing data was also consistent with the testing results of transplanted embryo.

Conclusions: The novel mutation of c. 156 G>T in CD40LG gene probably leads to XHIGM by nonsense-meditated mRNA decay (NMD), and complex PGT of preimplantation genetic testing for monogenic disease (PGT-M), aneuploidy (PGT-A), structural rearrangement (PGT-SR), and HLA-matching (PGT-HLA) can be performed in pedigree with both X-linked hyper IgM syndrome and Robertsonian translocation.

Keywords: CD40LG; HLA; PGT; Robertsonian translocation; XHIGM.

MeSH terms

  • Aneuploidy
  • Biopsy
  • Blastocyst / metabolism
  • CD40 Ligand / genetics*
  • Female
  • Fertilization in Vitro
  • Genetic Testing / trends
  • HLA Antigens / genetics*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Hyper-IgM Immunodeficiency Syndrome, Type 1 / diagnosis*
  • Hyper-IgM Immunodeficiency Syndrome, Type 1 / genetics
  • Hyper-IgM Immunodeficiency Syndrome, Type 1 / pathology
  • Pregnancy
  • Preimplantation Diagnosis*
  • Sperm Injections, Intracytoplasmic
  • Translocation, Genetic / genetics


  • HLA Antigens
  • CD40 Ligand