Detection of ASXL1 Codon 646 Variant Using Amplicon-Based Next-Generation Sequencing

• DOI: 10.15263/jlmqa.2022.44.2.76
• Publication Date: 2022-06-30T08:58:14Z
• Authors: Miyoung Kim; Nan Young Kim; Sangkyoon Hong; Jiwon Lee; Yonggeun Cho; Han-Sung Kim; Hee Jung Kang; Young Kyung Lee
• Keywords: Amplicon

Background:The ASXL1 codon 646 variant is the most common that negatively impacts prognoses of patients with myeloid malignancies, particularly those myelodysplastic syndromes and acute leukemia.However, it has been suggested this mutation not somatic but rather an artifact next-generation sequencing (NGS) owing to its location in 8 bp guanine mononucleotide repeat.In study, we evaluated performance amplicon-based NGS discriminating variant.Methods: Amplicon-based was performed on Myeloid DNA Reference Standard HD829 varying reference material dilution ratios using TruSight panel a MiSeqDx system.Results: The expected measured allele frequencies (VAFs) were 40.00% 18.65%, respectively.The VAFs materials serially diluted at 1:1, 1:2, 1:4, 1:8 9.09%, 5.82%, 1.92%, 2.87%, respectively (y=0.4391x+0.8642;r 2 =0.9846).Most other variants showed comparable VAFs. Conclusions:The approximately half their values, suggesting difficulties correct detection NGS.