AUTHOR=Lan Yueyun , Zhou Hong , He Sheng , Shu Jinhui , Liang Lifang , Wei Hongwei , Luo Jingsi , Wang Caizhu , Zhao Xin , Qiu Qingming , Huang Peng 

TITLE=Appropriate whole genome amplification and pathogenic loci detection can improve the accuracy of preimplantation genetic diagnosis for deletional α-thalassemia

JOURNAL=Frontiers in Endocrinology

VOLUME=14

YEAR=2024

URL=https://www.frontiersin.org/journals/endocrinology/articles/10.3389/fendo.2023.1176063

DOI=10.3389/fendo.2023.1176063

ISSN=1664-2392

ABSTRACT=<sec><title>Objective</title><p>To improve the accuracy of preimplantation genetic testing (PGT) in deletional α-thalassemia patients.</p></sec><sec><title>Design</title><p>Article.</p></sec><sec><title>Patient(s)</title><p>fifty-two deletional α-thalassemia couples.</p></sec><sec><title>Intervention(s)</title><p>Whole genome amplification (WGA), Next-generation sequencing (NGS) and PCR mutation loci detection.</p></sec><sec><title>Main outcome measures</title><p>WGA, Single nucleotide polymorphism (SNP) and PCR mutation loci detection results; Analysis of embryo chromosome copy number variation (CNV).</p></sec><sec><title>Results</title><p>Multiple Displacement Amplification (MDA) and Multiple Annealing and Looping–Based Amplification Cycles (MALBAC) methods for PGT for deletional α-thalassemia. Blastocyst biopsy samples (n = 253) were obtained from 52 deletional α-thalassemia couples. The results of the comparison of experimental data between groups MALBAC and MDA are as follows: (i) The average allele drop-out (ADO) rate, MALBAC <italic>vs.</italic> MDA = 2.27% ± 3.57% <italic>vs.</italic> 0.97% ± 1.4%, <italic>P</italic>=0.451); (ii) WGA success rate, MALBAC <italic>vs.</italic> MDA = 98.61% <italic>vs.</italic> 98.89%, <italic>P</italic>=0.851; (iii) SNP haplotype success rate, MALBAC <italic>vs.</italic> MDA = 94.44% <italic>vs.</italic> 96.68%, <italic>P</italic>=0.409; (iv) The result of SNP haplotype analysis is consistent with that of Gap-PCR/Sanger sequencing results, MALBAC <italic>vs.</italic> MDA = 36(36/72, 50%) <italic>vs.</italic> 151(151/181, 83.43%), <italic>P</italic>=0; (v) Valid SNP loci, MALBAC <italic>vs.</italic> MDA = 30 ± 9 <italic>vs.</italic> 34 ± 10, <italic>P</italic>=0.02; (vi) The mean CV values, MALBAC <italic>vs.</italic> MDA = 0.12 ± 0.263 <italic>vs.</italic> 0.09 ± 0.40, <italic>P</italic>=0.916; (vii) The average number of raw reads, MALBAC <italic>vs.</italic> MDA =3244259 ± 999124 <italic>vs.</italic> 3713146 ± 1028721, <italic>P</italic>=0; (viii) The coverage of genome (%), MALBAC <italic>vs.</italic> MDA = 5.02 ± 1.09 <italic>vs.</italic> 5.55 ± 1.49, <italic>P</italic>=0.008.</p></sec><sec><title>Conclusions</title><p>Our findings indicate that MDA is superior to MALBAC for PGT of deletional α-thalassemia. Furthermore, SNP haplotype analysis combined with PCR loci detection can improve the accuracy and detection rate of deletional α-thalassemia.</p></sec>