b'Next-Generation Sequencing (NGS) | CE-IVD AssaysIGH Assay Next-Generation Sequencing (NGS) | CE-IVD AssaysLymphoTrack Dx IGH FR1/2/3 AssaysAssay Description regions (FR1, FR2, or FR3) within the V Hand the J Hregions described LymphoTrack Dx IGH FR1 Assaysin lymphoid malignancies.Targeting all three framework regions The LymphoTrack Dx IGH FR1 Assay for the Illumina MiSeq or Thermosignificantly reduces the risk of not being able to detect the presence of clonality, as somatic hypermutations in the primer binding sites of Fisher Scientific Ion S5 TMand Ion PGM TMis an in vitro diagnostic productthe involved VH gene segments can impede DNA amplification. 1 The intended for next-generation sequencing (NGS) based determinationincluded primers are designed with Illumina or Thermo Fisher Scientific of the frequency distribution of IGH gene rearrangements as well asadapters and indices (8-24 and 12, respectively). This allows up to 24 the degree of somatic hypermutation of rearranged genes in patientssamples on MiSeq and 12 samples on Ion PGM and Ion S5 to be suspected with having lymphoproliferative disease.This assay aids in thesequenced at the same time with any of the individual FRs. identification of lymphoproliferative disorders as well as providing an aid in determining disease prognosis. In addition, amplicons generated with different FR master mixes or LymphoTrack Dx IGH FR1/2/3 AssaysInvivoscribe LymphoTrack Dx kits (such as IGK or TRG) can be pooled The LymphoTrack Dx IGH FR1 Assay for the Illumina MiSeq or Thermotogether in the same sequencing library to reduce testing costs. The Fisher Scientific Ion S5 TMand Ion PGM TMis an in vitro diagnostic productassociated LymphoTrack Dx Software provides interpretation of the data intended for next-generation sequencing (NGS) based determinationvia a simple and streamlined method of analysis and visualization.of the frequency distribution of IGH gene rearrangements as well asBy following the guidelines provided in the instructions for use, samples the degree of somatic hypermutation of rearranged genes in patientscan be interpreted for evidence of clonality suspected with having lymphoproliferative disease.This assay aids in theand SHM rates.identification of lymphoproliferative disorders as well as providing an aidPositive clonal (SHM negative) and negative polyclonal DNA controls in determining disease prognosis. are included in kits. A clonal SHM positive control can be purchased separately (cat#: 4-088-0008).This LymphoTrack Dx IGH FR2 Assay is an in vitro diagnostic product intended for next-generation sequencing (NGS) for the Illumina MiSeqBackgroundor Thermo Fisher Scientific Ion S5 TMand Ion PGM TMinstruments. The assay will determine the frequency distribution of IGH V H -J HgeneThe human immunoglobulin heavy chain (IGH) gene locus on rearrangements in patients suspected with having lymphoproliferativechromosome 14 (14q32.3) includes 46-52 functional and 30 non-disease.This assay aids in the identification of lymphoproliferativefunctional variable (V H ), 27 functional diversity (D H ), and 6 functional disorders. joining (J H ) gene segments. The V Hgene segments can be further broken down into three conserved frameworks (FR) and three variable The LymphoTrack Dx IGH FR3 Assay is an in vitro diagnostic productcomplementarity-determining regions (CDRs).intended for next-generation sequencing (NGS) for the Illumina MiSeq or Thermo Fisher Scientific Ion S5 TMand Ion PGM TMinstruments.TheDuring development of lymphoid cells, antigen receptor genes undergo assay will determine the frequency distribution of IGH V H -J Hgenesomatic gene rearrangements. 2Specifically during B-cell development, rearrangements in patients suspected with having lymphoproliferativeIGH molecules are assembled from multiple polymorphic gene segments disease.This assay aids in the identification of lymphoproliferativethat undergo rearrangements generating V H -D H -J Hcombinations unique disorders. in both length and sequence. 3Since leukemias and lymphomas originate from the malignant transformation of individual lymphoid cells, all Summary and Explanation of the Test leukemias and lymphomas generally share one or more cell-specific or clonal antigen receptor gene rearrangements.The LymphoTrack Dx IGH Assays represent a significant improvement over conventional clonality assessment methods utilizing fragmentIn addition, the IGHV hypermutation status obtained with the analysis by providing four important and complementary uses LymphoTrack Dx IGH FR1 master mixes, provides important prognostic in a single workflow: information for patients with chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL). The SHM rate has been shown 1.Detection of clonal populations. to have clinical relevance for CLL, as there is a clear distinction in the 2. Identification of sequence information and gene segment median survival of patients with and without SHM. 4 utilization.Specimen Requirement3. The LymphoTrack Dx IGH framework 1 FR1 master mixes provide thedegree of SHM in the immunoglobulin variable heavy chain (IGHV)50 ng of high-quality genomic DNA.gene locus.4.The ability to track sequences in subsequent samples with theReferencesInvivoscribe LymphoTrack MRD* Software. For more information,1. Evans, P. A. et al., (2007). Leukemia 21, 207-14. please refer to page 54 and 55.2. Tonegawa, S. (1983). Nature 302, 575-581. 3. Miller JE. (2013) Molecular Genetic Pathology (2nd Edition.,sectionsThese assays utilize a single multiplex master mix to target each30.2.7.13 and 30.2.7.18). conserved IGH Framework Region (FR1, FR2, and FR3) within the V Hand4. Ghia et al., Blood 105:1678-1685 (2005).the J Hregions described in lymphoid malignancies. Each singlemultiplex master mix targets one of the conserved IGH framework 28*MRD Software can be used to track sequences generated by either LymphoTrack Assays - MiSeq or Ion S5/PGM. MRD applications are for Research Use Only. To obtain a copy, please contact your local distributor or send an e-mail to [email protected]'