Long-Read Amplicon Sequencing for the Detection of TERT Promoter Variant Clonal Hematopoiesis in Patients with Telomere Biology Disorders

Acquired TERT promoter (TERTp) variants are found in the blood of patients with telomere biology disorders and carry diagnostic and prognostic significance. Detection of these variants is challenging because of low variant allele frequencies (VAFs) and high GC content. The sensitivity of long-read amplicon sequencing with deepSNV analysis, referred to as LR-deep AmpSeq, for TERTp variant detection was tested. Among 47 patients with telomere biology disorder features, an average depth of coverage of 7943× was achieved, and seven TERTp variants were detected in six individuals (13%) with VAFs ranging from 0.006 to 0.33. These results demonstrate that LR-deep AmpSeq is a sensitive, cost-effective method to detect low VAF TERTp variants.