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M. Zaliova et al.
data (analysis of hyper/hypo-diploidy, analysis of dele- tions involved in IKZF1plus pattern), the genomic PCR for the ERGdel detection represents an “extra” method and, moreover, it suffers from several disadvantages (as men- tioned above) which may discourage diagnostic centers from introducing it. Here we present a novel method, AmpliSeq, which is comparably sensitive to original genomic PCR and overcomes the disadvantages of Sanger sequencing of PCR products. Massive parallel sequencing (MPS) becomes a standard technology, and is used in
many routine diagnostic laboratories. The list of its appli- cations is still growing, from whole genome/exome/tran- scriptome sequencing to custom panels including, e.g. screening of immunoreceptor genes [immunoglobulin (IG) and T-cell receptor (TR)] rearrangements used for the iden- tification of targets for MRD monitoring. AmpliSeq for ERGdel detection can be easily coupled with such screen- ing of IG/TR rearrangements or with other MPS applica- tions. Moreover, as the analysis of IG/TR and ERGdel can use analogous algorithms, development of common ana-
Figure 5. Schematic representation of the IntERGdel repertoire in 24 patients with ERGdel-positivity by amplicon sequencing. Distinct, manually curated IntERGdel clones are shown as bars colored according to the 3’ breakpoint site cluster (BP) used. Width of bars corresponds to the relative size (number of reads) of individual clones. SNP: single nucleotide polymorphism; PCR: polymerase chain reaction; UPN: unique patient number.
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haematologica | 2019; 104(7)