Non-Invasive β-Thalassemia Diagnosis Using Cell FreeDNA Recovered from Blas tocele Fluid and Spent CultureMedium from Pre-Implantation Human Embryo

Background: To overcome the limitations of invasive embryobiopsy, a non-invasive alternative to preimplantation genetictes ting (PGT) has been recently introduced. This s tudy aimedto inves tigate the efficacy of cell-free DNA (CF-DNA) in thespent culture medium combined with blas tocoel fluid (ECB)versus blas tomere biopsy for β-thalassemia Diagnosis.Materials and Methods: This s tudy involved data from ۸ couples.A total of ۲۲ samples of ECB from fresh group who underwentPGT for β -thalassemia and ۶ ECB samples from donatedfrozen group and ۱۰ control media were collected and analyzed.Results: In fresh group, the HBB mutation analysis and haplotypingresults of ۹ of ۲۲ samples (۴۰.۹%) were concordant withthe biopsy results. The concordance rate in the donated frozengroup was higher than that in fresh group (۴/۶, ۶۶.۶%; P<۰.۰۵).۷۵% of markers were semi-informative. Marker failure ratewere significantly higher in fresh group (۳۴.۵% vs ۴.۸%;P<۰.۰۵). No significant relationship between allele dropout andfail number of all markers and embryo morphology was seen(P>۰.۰۵). Kappa- Agreement value between CF-DNA isolatedfrom culture media and blas tomere biopsy was ۰.۵۱۶ which ismoderate. The specificity, sensitivity, positive predictive value,and negative predictive value of CF-DNA to detect HBB mutationwere ۶۷, ۱۰۰, ۱۰۰, and ۶۷, respectively.Conclusion: We found that ECB for detecting β-thalassemia in donatedfrozen group was better than ECB in fresh group. Advancesin DNA extraction, amplification technique, and tes ting may allowfor PGT as a non-invasive approach without biopsy in the future.

کلیدواژه ها:

Beta-Thalassemia ، Blas tocys t ، Culture Media ، MicrosatelliteRepeats ، Preimplantation Diagnosis