b'Next-Generation Sequencing (NGS) | CE-IVD AssaysIGHV Leader Somatic Hypermutation Assay Next-Generation Sequencing (NGS) | CE-IVD Assays IGHV Leader Somatic Hypermutation Assay Next-Generation Sequencing (NGS) | CE-IVD AssaysLymphoTrack Dx IGHV Leader Somatic Hypermutation Assay Reagents - MiSeqDetectionKit A Components Panel Components(includes all master mixes from Kit A plus the items below)Master Mix Name Index # Master Mix Name Index # Master Mix Name Index #Assay Description Background IGH Leader MiSeq 01 A001 IGH Leader MiSeq 09 A009 IGH Leader MiSeq 18 A018IGH Leader MiSeq 02 A002 IGH Leader MiSeq 10 A010 IGH Leader MiSeq 19A019The LymphoTrack Dx IGHV Leader Somatic Hypermutation AssayThe human immunoglobulin heavy chain (IGH) gene locus on for the IlluminaMiSeqis an in vitro diagnostic product intendedchromosome 14 (14q32.3) includes 46-52 functional and 30IGH Leader MiSeq 03 A003 IGH Leader MiSeq 11 A011 IGH Leader MiSeq 20 A020for next-generation sequencing (NGS) based determination of thenonfunctional variable (V H ) gene segments, 27 functional diversityIGH Leader MiSeq 04 A004 IGH Leader MiSeq 12 A012 IGH Leader MiSeq 21 A021frequency distribution of IGH gene rearrangements, as well as the(D H ) gene segments, and 6 functional joining (J H ) gene segmentsIGH Leader MiSeq 05 A005 IGH Leader MiSeq 13 A013 IGH Leader MiSeq 22 A022degree of somatic hypermutation (SHM) of rearranged genes inspread over 1,250 kilobases.patients suspected of having lymphoproliferative disease. This assayIGH Leader MiSeq 06 A006 IGH Leader MiSeq 14 A014 IGH Leader MiSeq 23 A023aids in the identification of lymphoproliferative disorders as well asDuring B-cell development, genes encoding the IGH protein areIGH Leader MiSeq 07 A007 IGH Leader MiSeq 15 A015 IGH Leader MiSeq 25 A025providing an aid in determining disease prognosis. If you would likeassembled from multiple polymorphic gene segments that undergo to test for IGHV somatic hypermutation using the Thermo Fisher Ionrearrangements and selection, generating cell specific V H -D H -J H IGH Leader MiSeq 08 A008 IGH Leader MiSeq 16 A016 IGH Leader MiSeq 27 A027PGM or Ion S5 platform, please refer to the LymphoTrack Dx IGH FR1rearrangements that are unique in both length and sequence. 1 Controls ControlsAssay (9-121-0007).Since leukemias and lymphomas originate from the malignantIGH SHM POS (+) Qty. 1 IGH SHM POS (+) Qty. 3transformation of individual lymphoid cells, all leukemias and Summary and Explanation of the Test lymphomas generally share one or more cell-specific or "clonal"IGH POS (+) Qty. 1 IGH POS (+) Qty. 3The NGS LymphoTrack Dx IGHV Leader Somatic Hypermutationantigen receptor gene rearrangements. Therefore, tests that detectNGS NEG (-) Qty. 1 NGS NEG (-) Qty. 3Assay for the Illumina MiSeq represents a significant improvementIGH clonal rearrangements can be useful in the study of B- cell over clonality assays using fragment analysis as it efficiently detectsmalignancies. An additional level of diversity is further generated in the majority of IGH gene rearrangements using a single multiplexthe antigen receptors by introducing point mutations in the variableMerge% TotalCumulative Mutation rate In-frameNo stop regions, also named SHM. In instances where there is a high degreeRank Sequence Length count V-gene J-gene reads %partial V-gene(Y/N) codon V-coveragemaster mix, identifies the DNA sequence specific for each clonal geneof SHM, there is the risk that primers located within the variable region(%) (Y/N)rearrangement, and assesses the somatic hypermutation rate ofwill not be able to bind and clonal products will not amplify. In these clonal samples in the same workflow. cases, the leader primers located upstream of the variable region1 TTCTCGTGGTG 455 29603 IGHV4-59_08 IGHJ4_02 9.93 9.93 11.26 Y Y 98.63The single multiplex master mixes target the Leader (V H L) and thecan be beneficial for the detection of clonal products, due to the2 CTCGCCCTCCT 463 205 IGHV5-51_01 IGHJ4_02 0.07 9.99 0.00 Y Y 99.66joining (J H ) gene regions of the IGH locus and are designed withconserved nature of the V H L region. In addition, the SHM rate of the3 GGTTTTCCTTG 484 201 IGHV3-7_01 IGHJ4_02 0.07 10.06 7.77 Y Y 100.00Illumina adapters and indices (8 included in Kit A and 24 includedentire variable gene can be determined using the V H L primers.in the Panel). This allows for a one-step PCR reaction and pooling of4 CTCGCCCTCCT 463 185 IGHV5-51_01 IGHJ5_02 0.06 10.12 6.08 Y Y 99.32amplicons from several different samples and targets into a singleDetermining the immunoglobulin variable heavy chain gene (IGHV)5 CTCGCCCTCCT 469 170 IGHV5-51_01 IGHJ4_02 0.06 10.18 0.00 Y Y 99.32 hypermutation rate is considered a gold standard for determining the Illumina MiSeqrun. No post-PCR ligation step is required. prognosis of patients with chronic lymphocytic leukemia (CLL) 2and6 CTCGCCCTCCT 466 160 IGHV5-51_01 IGHJ4_02 0.05 10.23 0.00 Y Y 99.66The included LymphoTrack Dx Bioinformatics Software enablessmall lymphocytic lymphoma (SLL). In addition, NGS methods can7 CTGCTGCTGAC 460 159 IGHV2-5_10 IGHJ5_02 0.05 10.29 8.08 Y Y 97.64simplified analysis and visualization of individual sample data.improve disease stratification.8 GGTTTTCCTTG 493 156 IGHV3-48_02 IGHJ6_02 0.05 10.34 3.72 Y Y 98.99Positive (clonal positive, SHM negative), negative (polyclonal) and9 CTCGCCCTCCT 334 153 IGHV5-51_02 IGHJ2_01 0.05 10.39 3.72 Y N 27.70SHM (clonal positive, SHM positive) DNA controls are included in the kits. Specimen Requirement 10 CTCGCCCTCCT 334 152 IGHV5-51_02 IGHJ2_01 0.05 10.44 3.38 Y N 26.0150 ng of high-quality genomic DNA. Example Data. The top 10 sequences from a read summary generated by the LymphoTrack Dx Software - MiSeq with the SHM mutation rate and predictions pertaining to whether a sequence is in-frame or contains a premature stop codon are depicted. To learn more about the LymphoTrack Dx software, please refer to the LymphoTrack Dx Bioinformatics Software section.References1.Miller et al., Molecular Genetic Pathology (2nd ed.). Springer Ordering InformationScience & Business Media. 2013: 302.2.7.13 and 30.2.7.18.Catalog # Products Quantity Components2. Ghia et al., Leukemia 21: 2-3 (2007).9-121-0059 LymphoTrackDx IGHV Leader Somatic Hypermutation Assay Kit A - MiSeq 8 indices - 5 sequencing reactions each9-121-0069 LymphoTrackDx IGHV Leader Somatic Hypermutation Assay Panel - MiSeq 24 indices - 5 sequencing reactions each9-500-0009 LymphoTrackDx Software - MiSeq 1 CD complimentary with purchaseV H L V H 1 V H 2 V H 3 V H 4 V H 5 V H 6 V H 7 J H These products are CE-IVD assays for in vitro diagnostic use.Simplified representation of the immunoglobulin heavy chain (IGH) gene locus on chromosome 14. Depicted are the variable (V H ) and downstream consensus joining (J H ) region genes involved in rearrangements. Upstream of the variable gene segments, the leader sequence (V H L) is also depicted. Diversity region genes are not depicted.24Invivoscribe 2021|25'