Molecular

LymphoTrack IGH Assay

This research use only (RUO) assay identifies clonal IGH V-J rearrangements, the associated V-J region DNA sequences, and provides the frequency distribution of V region and J region segment utilization and the degree of somatic hypermutation of rearranged genes using the Illumina MiSeq® or Thermo Fisher / Life Technologies Ion PGM®.

The LymphoTrack® IGH FR1 assay represents a significant improvement over existing clonality assays as it efficiently detects the majority of IGH gene rearrangements using a single multiplex master mix that identifies the DNA sequence specific for each clonal gene rearrangement, and determines the degree of somatic hypermutation of rearranged genes.

The LymphoTrack® IGH FR1 assay represents a significant improvement over existing fragment analysis clonality assays by providing three important and complementary uses:
– Detects initial clonal populations
– Identifies sequence information required to track clonal rearrangements in subsequent samples
– Provides detailed sequence information on the degree of somatic hypermutation (SHM) of the immunoglobulin variable heavy chain (IGHV) gene

Primers included in the master mix are designed with Illumina® adapters and indices (8-24) or Thermo Fisher / Life Technologies® adaptors and indices (12). This allows for a one-step PCR reaction and pooling of amplicons from several different samples for loading on the Illumina MiSeq® flow-cell or Ion PGM® chips.

LymphoTrack assays represent a significant improvement over existing clonality assays using fragment analysis as they efficiently detect the majority of IGH and TRG gene rearrangements using a single multiplex master mix for each locus. At the same time, the assay identifies the DNA sequence specific to each clonal gene rearrangement.

The LymphoTrack® bioinformatics software enables simplified analysis and visualization of data generated from this assay.

Positive clonal and negative polyclonal DNA controls are included in the kit.
– LymphoTrack IGH Assay – MiSeq
– LymphoTrack IGH Assay – PGM

LymphoTrack IGH Somatic Hypermutation

This research use only (RUO) assay identifies clonal IGH V-J rearrangements, the associated V-J region DNA sequences, provides the frequency distribution of V region and J region segment utilization using the Illumina MiSeq® or Thermo Fisher / Life Technologies Ion PGM® platforms and defines the extent of somatic hypermutation present in the IGHV gene of analyzed samples.

The LymphoTrack® IGH Somatic Hypermutation (SHM) Assay for the Next-Generation Sequencing represents a significant improvement over clonality assays using fragment analysis as it efficiently detects the majority of IGH gene rearrangements using a single multiplex master mix and identifies the DNA sequence specific for each clonal gene rearrangement. Therefore, this product aids in the detection of initial clonal populations, identifies sequence information required to track those clones in subsequent samples, and determines the somatic hypermutation (SHM) status of samples.

The master mix included in the MiSeq® version of this assay targets the Leader (VHL) and the joining (J) gene regions of IGH and are designed with Illumina® adapters and indices (8 included in Kits A, B, or C and 24 included in the Panel). The PGM® version of the SHM assay includes forward primers that target the conserved framework region 1 (FR1) and the J region of IGH and are designed with PGM®. IonExpress indices (12). Including adapters and indices in the primer design allows for a one-step PCR followed by directly pooling of amplicons from several different samples for sequencing on a Illumina MiSeq® flow-cell or Ion PGM® chip.

The included LymphoTrack® Bioinformatics software enables simplified analysis and visualization of data generated from this assay.

Positive, negative, and SHM controls are included in the kit.
– LymphoTrack IGH Somatic Hypermutation Assay – MiSeq
– LymphoTrack IGH Somatic Hypermutation Assay – PGM

LymphoTrack IGK Assay

This research use only (RUO) assay identifies clonal IGK Vк-Jк, Vк-Kde, and intron-Kde (INTR – Kde) rearrangements, the DNA sequences of these rearrangements, and provides the distribution frequency of Vк and Jк, Vк-Kde, and INTR – Kde segment utilization using the Illumina MiSeq® or Thermo Fisher / Life Technologies Ion PGM® platforms.

This assay utilizes a single multiplex master mix to target conserved V and J regions of IGK that are described in lymphoid malignancies. Primers are designed with Illumina® adapters or Thermo Fisher / Life Technologies® adapters and up to 24 different indices; thereby allowing amplicons generated from different IGK master mixes to be pooled together to generate a library for loading onto a single MiSeq® flow cell or Ion PGM® chip for sequencing. The associated LymphoTrack® Software provides interpretation of the data via a simple and streamlined method of analysis and visualization of data. By following the guidelines provided in the Instructions for Use, samples can be easily interpreted for the evidence or no evidence of clonality.
– LymphoTrack IGK Assay – MiSeq
– LymphoTrack IGK Assay – PGM

LymphoTrack TRG Assay

This research use only assay identifies clonal TRG V-J rearrangements, the associated V-J region DNA sequences, and provides the frequency distribution of V region and J region segment utilization using the Illumina MiSeq® or Thermo Fisher / Life Technologies Ion PGM®. This LymphoTrack® TRG assay represents a significant improvement over existing clonality assays as it efficiently detects the vast majority of TRG gene rearrangements using a single multiplex master mix and, at the same time, the assay identifies the DNA sequence specific for each clonal gene rearrangement.

Therefore, this product has two important and complementary uses: it both aids in the detection of initial clonal populations, and identifies sequence information required to track those clones in subsequent samples. Our single multiplex master mix targets all conserved regions within the variable (V) and the joining (J) region genes described in lymphoid malignancies. This is critical for comprehensive analysis of samples, as some T-cell lymphoproliferative disorders involve V and J segments that would not be identified with existing assays or Vγ and Jγ primer sets. The average size of the TRG gene rearrangement PCR amplicons generated using this assay was designed to be compatible with testing fragmented DNA isolated from more challenging samples (e.g., FFPE sections). Positive and negative DNA controls are included in the kit.

For more information, please visit the IVS Website.