Development and validation of a fetal genotyping assay with potential for noninvasive prenatal diagnosis of hereditary hearing loss

Prenat Diagn. 2016 Dec;36(13):1233-1241. doi: 10.1002/pd.4962. Epub 2016 Dec 9.

Abstract

Objective: Inherited non-syndromic hearing loss (NSHL) is a common sensory disorder that afflicts otherwise healthy individuals. The aim of the study was to evaluate the performance of circulating single molecule amplification and re-sequencing technology (cSMART) for non-invasive prenatal testing (NIPT) of NSHL.

Method: Neonatal inheritance of NSHL mutations was determined from bloodspots using SNaPshot genotyping. NIPT of cell-free DNA for fetal NSHL mutations in the GJB2, GJB3 and SLC26A4 genes was performed by a multiplex cSMART assay. The percentage of mutant alleles was used to deduce fetal DNA fractions and assign fetal genotypes.

Results: A total of 25 plasma samples selected with different fetal NSHL genotypes were coded and retrospectively analyzed by NIPT. Three normal fetuses, 18 carrier fetuses comprising seven GJB2 109G>A, four GBJ2 235delC, three GJB2 299-300delAT and four SLC26A4 IVS7-2A>G heterozygotes and four affected fetuses comprising two GJB2 109G>A homozygotes, one GBJ2 235delC homozygote and one compound GJB2 235delC/299-300delAT heterozygote were identified. All 25 fetal genotypes determined by the cSMART assay were concordant with neonatal genotypes.

Conclusion: The cSMART assay applied to cell-free DNA isolated from maternal plasma of pregnant women is highly accurate for calling correct fetal NSHL genotypes. © 2016 John Wiley & Sons, Ltd.

Publication types

  • Validation Study

MeSH terms

  • Connexin 26 / genetics
  • Connexins / genetics
  • DNA / blood
  • Female
  • Genetic Testing / methods
  • Genotype*
  • Genotyping Techniques / methods*
  • Hearing Loss / congenital*
  • Hearing Loss / genetics*
  • Heterozygote
  • Homozygote
  • Humans
  • Infant, Newborn
  • Membrane Transport Proteins / genetics
  • Mutation
  • Phenotype
  • Pregnancy
  • Prenatal Diagnosis / methods*
  • Sulfate Transporters

Substances

  • Connexins
  • Membrane Transport Proteins
  • SLC26A4 protein, human
  • Sulfate Transporters
  • Connexin 26
  • GJB3 protein, human
  • DNA