Interpretation Turn-around Time Specimen Requirements Shipping Conditions Storage Conditions  An interpretive report will be issued indicating whether evidence of clonality was detected. The report further provides a summary of the top 5 merged sequences, including the % total reads, the rearrangement class and the sequence. 5 to 10 business days •  3 mL of peripheral blood in Heparin, EDTA or ACD •  1 mL of bone marrow in Heparin, EDTA or ACD •  500 ng of previously isolated DNA • FFPE tissue samples Ambient or Cool; Do not freeze • Room Temp up to 72 hours •  2-8 °C up to 7 days Test Name IGH FR1 clonality assay IGH FR2 clonality assay IGH FR3 clonality assay IGHV Leader Somatic Hypermutation Clonality Assay Assay Type Next-Generation Sequencing (NGS) This test is performed by using the LymphoTrack® Assay from Invivoscribe Method Description For detection of the vast majority of clonal IGH VH-JH rearrangements, including the associated VH-JH region DNA sequences, a multiplex master mix targeting the conserved framework region 1, framework region 2, or framework region 3, as well as the joining region, is used for PCR amplification. Next-generation sequencing of the PCR products is used to identify DNA sequences specific to clonal gene rearrangements. Bioinformatics tools facilitate the characterization of sequences present at greater than 2.5% or 5% of the population. These sequences can be used to track specific clonal populations. Indications for Testing •  Identify clonality in atypical lymphoproliferative disorders •  Support a differential diagnosis between reactive lesions and hematologic malignancies •  Assign presumptive lineage in mature monoclonal lymphoproliferative disorders  • Monitor and evaluate disease recurrence Clonaltiy Tests LabPMM Services Catalog 2019 | 31 Clinical Information Lymphoid cells are different from the other somatic cells in the body as during development, the antigen receptor genes of these cells undergo somatic gene rearrangement1 . The human immunoglobulin heavy chain (IGH) gene locus on chromosome 14 (14q32.3) includes 46-52 functional and 30 non-functional variable (VH) gene segments, 27 functional diversity (DH) gene segments, and 6 functional joining (JH) gene segments spread over 1250 kilobases. During B-cell development, genes encoding the IGH molecules are assembled from multiple polymorphic gene segments that undergo rearrangements and selection. These gene rearrangements of the variable, diversity and joining segments generate VH-DH-JH combinations of unique length and sequence for each cell2-3 . Since leukemia and lymphomas originate from the malignant transformation of individual lymphoid cells, all leukemias and lymphomas generally share one or more cell-specific or “clonal” antigen receptor gene rearrangements. Clonality does not always imply malignancy; all results must be interpreted in the context of all of the other available diagnostic criteria. Tests that detect IGH clonal rearrangements are useful in the characterization, monitoring, and treatment of B- and T-cell malignancies. IGH Clonality Assays References 1. Tonegawa S (1983) Somatic Generation of Antibody Diversity. Nature 302:575-581. 2.  Trainor KJ et al. (1990). Monoclonality in B-lymphoproliferative disorders detected at the DNA level. Blood 75:2220-2222. 3. JE Miller et al., Molecular Genetic Pathology (2013, 2nd ed.) Springer Science & Business Media 302.2.7.13 and 30.2.7.18. 30