b"Next-Generation Sequencing (NGS) | CE-IVD AssaysIGH Assay Next-Generation Sequencing (NGS) | CE-IVD Assays IGH 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 describedSimplified Representation of the IGH GeneLymphoTrack Dx IGH FR1 Assaysin lymphoid malignancies.Targeting all three framework regionsSimplified depiction of variable (V H ) and downstream consensus The LymphoTrack Dx IGH FR1 Assay for the Illumina MiSeq or Thermosignificantly reduces the risk of not being able to detect the presenceD H J H joining (J H ) region genes involved in gene rearrangements.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 TheV L V H -FR1 V H -FR2 V H -FR3J Hprimers intended for next-generation sequencing (NGS) based determinationincluded primers are designed with Illumina or Thermo Fisher ScientificV Hfamily primersof 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 aidReagents - MiSeq Detectionin determining disease prognosis.In addition, amplicons generated with different FR master mixes or Invivoscribe LymphoTrack Dx kits (such as IGK or TRG) can be pooledThe LymphoTrack Dx IGH FR1/2/3 Assays contain components from respective individual FR kit A's or panels.LymphoTrack Dx IGH FR1/2/3 Assaystogether in the same sequencing library to reduce testing costs. The The LymphoTrack Dx IGH FR1 Assay for the Illumina MiSeq or Thermoassociated LymphoTrack Dx Software provides interpretation of the dataLymphoTrack Dx IGH FR1 Components LymphoTrack Dx IGH FR2 Components LymphoTrack Dx IGH FR3 ComponentsFisher Scientific Ion S5 TMand Ion PGM TMis an in vitro diagnostic productvia a simple and streamlined method of analysis and visualization. Master Mix Name Index # Master Mix Name Index # Master Mix Name Index #intended for next-generation sequencing (NGS) based determinationBy following the guidelines provided in the instructions for use, samples of the frequency distribution of IGH gene rearrangements as well ascan be interpreted for evidence of clonalityIGH FR1 MiSeq 01 A001 IGH FR2 MiSeq 01 A001 IGH FR3 MiSeq 01 A001the degree of somatic hypermutation of rearranged genes in patientsand SHM rates. IGH FR1 MiSeq 02 A002 IGH FR2 MiSeq 02 A002 IGH FR3 MiSeq 02 A002suspected with having lymphoproliferative disease.This assay aids in the identification of lymphoproliferative disorders as well as providing an aidPositive clonal (SHM negative) and negative polyclonal DNA controlsIGH FR1 MiSeq 03 A003 IGH FR2 MiSeq 03 A003 IGH FR3 MiSeq 03 A003in determining disease prognosis. are included in kits. A clonal SHM positive control can be purchasedIGH FR1 MiSeq 04 A004 IGH FR2 MiSeq 04 A004 IGH FR3 MiSeq 04 A004KIT A KIT A KIT Aseparately (cat#: 4-088-0008).This LymphoTrack Dx IGH FR2 Assay is an in vitro diagnostic productIGH FR1 MiSeq 05 A005 IGH FR2 MiSeq 05 A005 IGH FR3 MiSeq 05 A005intended for next-generation sequencing (NGS) for the Illumina MiSeqBackground IGH FR1 MiSeq 06 A006 IGH FR2 MiSeq 06 A006 IGH FR3 MiSeq 06 A006or 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 onIGH FR1 MiSeq 07 A007 IGH FR2 MiSeq 07 A007 IGH FR3 MiSeq 07 A007rearrangements in patients suspected with having lymphoproliferativechromosome 14 (14q32.3) includes 46-52 functional and 30 non- IGH FR1 MiSeq 08 A008 IGH FR2 MiSeq 08 A008 IGH FR3 MiSeq 08 A008disease.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 furtherIGH FR1 MiSeq 09 A009 IGH FR2 MiSeq 09 A009 IGH FR3 MiSeq 09 A009broken down into three conserved frameworks (FR) and three variableIGH FR1 MiSeq 10 A010 IGH FR2 MiSeq 10 A010 IGH FR3 MiSeq 10 A010The LymphoTrack Dx IGH FR3 Assay is an in vitro diagnostic productcomplementarity-determining regions (CDRs).intended for next-generation sequencing (NGS) for the Illumina MiSeqIGH FR1 MiSeq 11 A011 IGH FR2 MiSeq 11 A011 IGH FR3 MiSeq 11 A011or Thermo Fisher Scientific Ion S5 TMand Ion PGM TMinstruments.TheDuring development of lymphoid cells, antigen receptor genes undergoIGH FR1 MiSeq 12 A012 IGH FR2 MiSeq 12 A012 IGH FR3 MiSeq 12 A012PANEL PANEL PANELassay 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 segmentsIGH FR1 MiSeq 13 A013 IGH FR2 MiSeq 13 A013 IGH FR3 MiSeq 13 A013disease.This assay aids in the identification of lymphoproliferativethat undergo rearrangements generating V H -D H -J Hcombinations uniqueIGH FR1 MiSeq 14 A014 IGH FR2 MiSeq 14 A014 IGH FR3 MiSeq 14 A014disorders. in both length and sequence. 3Since leukemias and lymphomas originate from the malignant transformation of individual lymphoid cells, allIGH FR1 MiSeq 15 A015 IGH FR2 MiSeq 15 A015 IGH FR3 MiSeq 15 A015Summary and Explanation of the Test leukemias and lymphomas generally share one or more cell-specific orIGH FR1 MiSeq 16 A016 IGH FR2 MiSeq 16 A016 IGH FR3 MiSeq 16 A016clonal antigen receptor gene rearrangements.The LymphoTrack Dx IGH Assays represent a significant improvementIGH FR1 MiSeq 18 A018 IGH FR2 MiSeq 18 A018 IGH FR3 MiSeq 18 A018over conventional clonality assessment methods utilizing fragmentIn addition, the IGHV hypermutation status obtained with theIGH FR1 MiSeq 19 A019 IGH FR2 MiSeq 19 A019 IGH FR3 MiSeq 19 A019analysis 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) andIGH FR1 MiSeq 20 A020 IGH FR2 MiSeq 20 A020 IGH FR3 MiSeq 20 A020small lymphocytic lymphoma (SLL). The SHM rate has been shownIGH FR1 MiSeq 21 A021 IGH FR2 MiSeq 21 A021 IGH FR3 MiSeq 21 A0211.Detection of clonal populations. to have clinical relevance for CLL, as there is a clear distinction in the median survival of patients with and without SHM. 4IGH FR1 MiSeq 22 A022 IGH FR2 MiSeq 22 A022 IGH FR3 MiSeq 22 A0222. Identification of sequence information and gene segment IGH FR1 MiSeq 23 A023 IGH FR2 MiSeq 23 A023 IGH FR3 MiSeq 23 A023utilization. Specimen Requirement IGH FR1 MiSeq 25 A025 IGH FR2 MiSeq 25 A025 IGH FR3 MiSeq 25 A0253. 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. IGH FR1 MiSeq 27 A027 IGH FR2 MiSeq 27 A027 IGH FR3 MiSeq 27 A027gene locus. Kit A's contain indices IGH FRX A001 to A008. Panels contain all master mixes listed above.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.Controls in Individual Controls in Individual Controls in ComboControls in Comboplease refer to page 52 and 53.2. Tonegawa, S. (1983). Nature 302, 575-581.FR (1, 2, or 3) Kit A's FR (1, 2, or 3) Panels FR 1/2/3 Kit A FR 1/2/3 Panel3. 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).IGH POS (+) Qty. 1 IGH POS (+) Qty 3 IGH POS (+) Qty. 2 IGH POS (+) Qty. 6conserved IGH Framework Region (FR1, FR2, and FR3) within the V Hand4. Ghia et al., Blood 105:1678-1685 (2005). NGS NEG (-) Qty. 1 NGS NEG (-) Qty 3 NGS NEG (-) Qty. 2 NGS NEG (-) Qty. 6the J Hregions described in lymphoid malignancies. Each singlemultiplex master mix targets one of the conserved IGH framework 26*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 orInvivoscribe 2021|27send an e-mail to customerservice@invivoscribe.com"