b'Gel & Capillary | CE-IVD Assays IGH Gene Clonality AssaysGel and CapillaryAssaysIGH Gene Clonality AssaysAssay Description Performance Characteristics The IdentiClone IGH Gene Clonality Assays are in vitro diagnosticData from two independent studies that tested more than 300 products intended for PCR-based detection of clonal immunoglobulinpatient samples of varying types suggests the diagnostic accuracyheavy chain gene rearrangements in patients with suspectof selected IdentiClone tests to be 96%. In both peer-reviewed lymphoproliferations. studies, there were no clear false-positive results generated using the IdentiClone tests, and there was a high level of precision. 2Specifically, the IdentiClone IGH Gene Clonality Assays can be used to: The clinico-histopathological diagnosis correlated well with PCR results in a higher number of patients when compared withIdentify clonality in atypical lymphoproliferative disorders Southern Blot (SB) results, as seen below:PCR/SB concordance: 1Support a differential diagnosis between reactive lesions and hematologic malignancies IGH:93% sensitivity / 92% specificityAssign presumptive lineage in mature monoclonal IGK: 90% sensitivity / 90% specificity lymphoproliferative disorders IGL: 86% sensitivity / 92% specificity Identify tumor-specific markers (IGH gene rearrangements) TRB: 86% sensitivity / 98% specificity for post-treatment monitoring TRG: 89% sensitivity / 94% specificity Monitor and evaluate disease recurrence TRD: 83% sensitivity / 95% specificityPCR vs. SB analysis relative to histopathology and final diagnosis:Summary and Explanation of the TestPCR/SB concordance: 2PCR sensitivity: SB sensitivity: The Invivoscribe CE-marked IdentiClone Assays represent a uniqueIGH + IGK: 85% 98% 39% approach to PCR-based clonality testing. These standardized assaysTRB: 85% 96% 35%were carefully optimized, testing positive and negative controlReferencesamples using multiplex master mixes. Assay development was 1. JJM van Dongen et al., Leukemia 17:2257-2317 (2003). followed by extensive validation including the testing of more than 400 clinical samples using Revised European/American Lymphoma2. Y Sandberg et al., J. Mol. Diag. 7(4):495-503 (2005).(REAL) Classification. Testing was done at more than thirty prominent independent testing centers throughout Europe in a collaborative study known as the BIOMED-2 Concerted Action. Results from this BIOMED-2 study appear in Leukemia, a leading peer-reviewed journal.These test kits include six master mixes. The IGH Tube A, B, and C master mixes target the framework 1, 2, and 3 regions (respectively) within the variable (VH) region and the joining (JH) region of the immunoglobulin heavy chain locus. The IGH Tube D and E master mixes target the diversity and joining regions. The Specimen Control Size Ladder master mix targets multiple genes and generates a series of amplicons of approximately 100, 200, 300, 400, and 600 base pairs to ensure that the quality and quantity of input DNA is adequate to yield a valid result. A single thermal cycler program and similar detection methodologies are used with all of our gene clonalityFigure Legend: Simple representation of the organization of a rearranged assays. This improves consistency and facilitates cross-training on aimmunoglobulin heavy chain gene on chromosome 14q32.33. Black arrows broad range of different assays. represent the relative positions of primers that target the conserved framework (FR1-3) and diversity (D H 1-7) regions, and the downstream consensus J Hgene segments. The amplicon products generated from each of these regions can be differentially detected when fluorescent primer sets are used with capillary electrophoresis instruments that employ differential fluorescence detection.This assay is based on the EuroClonality/BIOMED-2 Concerted Action BMH4-CT98-3936.60'