Ann Lab Med.  2017 Jul;37(4):331-335. 10.3343/alm.2017.37.4.331.

Detection of Immunoglobulin Heavy Chain Gene Clonality by Next-Generation Sequencing for Minimal Residual Disease Monitoring in B-Lymphoblastic Leukemia

• Shin S
• Hwang IS
• Kim J
• Lee KA
• Lee ST
• Choi JR

• KMID: 2376791
• DOI: http://doi.org/10.3343/alm.2017.37.4.331

Abstract

Minimal residual disease (MRD) following B-lymphoblastic leukemia (B-ALL) treatment has gained prognostic importance. Clonal immunoglobulin heavy chain (IGH) gene rearrangement is a useful follow-up marker in B-ALL owing to its high positivity rate. We evaluated the performance and clinical applicability of a next-generation sequencing (NGS) assay for IGH rearrangement in B-ALL MRD monitoring. IGH rearrangement was tested by using fluorescence PCR-fragment analysis and the NGS assay in eight B-ALL patients. The NGS assay was run on two platforms: the Ion Torrent PGM (Thermo Fisher Scientific, USA) (18 samples from 1st to 7th patients) and the MiSeq system (Illumina, USA) (four samples from 8th patient). All initial diagnostic samples and four follow-up samples were positive for clonal IGH rearrangement with fluorescence PCR-fragment analysis and the NGS assay, and six follow-up samples were positive only with NGS. In one case with BCR-ABL1 translocation, BCR-ABL1 quantitative PCR was negative but the NGS IGH assay was positive just prior to full-blown relapse, suggesting the high sensitivity and clinical utility of the NGS assay. The NGS assay is proposed for MRD monitoring in B-ALL Additional studies are needed to confirm the clinical implications of cases showing positive results only in NGS.

Keyword

Immunoglobulin heavy chain gene rearrangement; Minimal residual disease; B-lymphoblastic leukemia; Next-generation sequencing

MeSH Terms

Fluorescence
Follow-Up Studies
Gene Rearrangement
Humans
Immunoglobulin Heavy Chains*
Immunoglobulins*
Leukemia*
Neoplasm, Residual*
Polymerase Chain Reaction
Recurrence
Immunoglobulin Heavy Chains
Immunoglobulins

Figure

• 

  Fig. 1 Next-generation sequencing (NGS) results using MiSeq (Illumina) for IGH clonality detection at diagnosis (A), follow-up (B, C), and full-blown relapse (D) in Patient 8. Each colored box represents the frequency of reads with an identical sequence. NGS showed a leukemic clone frequency of 57.7% (A) at initial diagnosis and of 14.3% (B) at 30 days after diagnosis (white arrows). (C) A residual leukemic clone frequency of 2.8% was found by NGS at two months before full-blown relapse, although PCR fragment analysis for IGH clonality detection and real-time PCR for BCR-ABL1 transcript quantification showed negative results.

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