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Test Code 5515 Immunoglobulin Heavy-Chain (IGH) Hypermutation (IGH V Region) Sequencing

Performing Laboratory

Barnes-Jewish Hospital Laboratory

Methodology

Extracted genomic DNA is analyzed by a laboratory-developed test using the LymphoTrack® Dx IGH Assay reagents from InVivoScribe Technologies. Massively parallel (“Next-generation”) sequencing of libraries is performed on a Life Technologies Ion Torrent S5™ sequencer. The LymphoTrack® Dx IGH clonality testing software is used to determine the IGHV gene rearrangement sequences, the relative proportion as a percentage of total sequencing reads, and mutation level. Limitations: The assay primers cannot detect 100% of possible IGHV rearrangements or mutations. Low DNA input, DNA degradation, and/or interfering substances may cause failure to detect an IGHV rearrangement by this assay.

 

Literature References

1. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983;302(5909):575-581.

2. Gazzola A, Mannu C, Rossi M, et al. The evolution of clonality testing in the diagnosis and monitoring of hematological malignancies. Therapeutic advances in hematology. 2014;5(2):35-47.

3. Parikh SA, Strati P, Tsang M, West CP, Shanafelt TD. Should IGHV status and FISH testing be performed in all CLL patients at diagnosis? A systematic review and meta-analysis. Blood. 2016;127(14):1752-1760. doi: 10.1182/blood-2015-10-620864

Specimen Requirements

Call Barnes-Jewish Molecular Diagnostics Laboratory at 314-454-8685 for collection instructions.

 

Forms: Molecular Diagnostics Laboratory Request for DNA Studies-Oncology in Special Instructions.

 

Submit only 1 of the following specimens:

 

Specimen Type: Blood

Container/Tube: Lavender top (EDTA)

Specimen Volume: Full tube

Specimen Minimum Volume: 1 to 2 mL

Collection Instructions:

1. Invert several times to mix blood. Clotted blood is not acceptable.

2. Specimen cannot be frozen.

 

Specimen Type: Bone marrow

Container/Tube: Lavender top (EDTA)

Specimen Volume: 1 to 2 mL

Collection Instructions:

1. Invert several times to mix bone marrow. Clotted bone marrow is not acceptable.

2. Specimen cannot be frozen.

 

 

Specimen Type: Tissue, formalin-fixed, paraffin-embedded

Container/Tube: Sterile, plastic microcentrifuge tube

Specimen Volume: Twenty, 10-micron sections of tissue

 

Specimen Type: Tissue, fresh

Container/Tube: Sterile, plastic container

Specimen Volume: 10 mm3

Additional Information: Forward frozen tissue on dry ice.

Day(s) Test Set Up

Monday

Turnaround Time:
STAT: not available
Routine: 7 to 10 business days

Reference Values

An interpretive report will be provided.

 

Test Classification and CPT Coding

81263 

Additional Information

For BJH Laboratory Use Only
Analyte Stability:

Blood is stable 0 to 4 hours at ambient temperature, 96 hours at 4° C.
FFPE tissue stable indefinitely.
Fresh-frozen tissue is stable at -20° C.

Laboratory Processing Instructions:
Test performed in BJH Molecular Diagnostics.

Specimen Transport Temperature

Ambient/Refrigerate NO/Frozen NO-Blood; Tissue, Formalin-Fixed, Paraffin-Embedded

Frozen on Dry Ice-Tissue, fresh; Separated Cell Pellets