Application of Cell-Free DNA Barcode-Enabled Single-Molecule Test for Non-Invasive Prenatal Testing of α-Thalassemia and β-Thalassemia.

PURPOSE: To assess the feasibility of using the cell-free DNA barcode-enabled single-molecule test (cfBEST) for non-invasive prenatal testing (NIPT) of α-thalassemia and β-thalassemia.

METHODS

Seventy two thalassemia carrier families participated in the study. The cfBEST method was employed to identify thalassemia genotypes in fetal cell-free DNA extracted from maternal plasma, targeting four α-thalassemia gene mutations and 13 common β-thalassemia gene mutations. Validation was conducted using gap-PCR and PCR-RDB through invasive prenatal diagnosis. All prenatal diagnosis results were later confirmed for accuracy.

RESULTS

cfBEST successfully identified 94.6% (88/93) of fetal alleles, achieving a sensitivity of 94% (95% CI, 83.45%-98.75%) and a specificity of 95.35% (95% CI, 84.19%-99.43%). All prenatal diagnosis results were followed up, and the follow-up findings were consistent with the prenatal diagnosis results.

CONCLUSION

The findings indicate that cfBEST is a reliable, precise, simple, and cost-efficient method suitable for non-invasive prenatal testing of α-thalassemia and β-thalassemia.