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B-cell IGK Gene Rearrangement by NGS

Test Code

M IGK

Test Synonyms

B-cell gene rearrangement, B-cell clonality

Associations

B-cell leukemia, lymphoma

Methodology

Next Generation Sequencing (NGS)

Turnaround Time

5-7 days

Specimen Requirements

5.0 mL (min. 3.0 mL) whole blood EDTA or Sodium Heparin
3.0 mL (min. 1.0 mL) bone marrow EDTA or Sodium Heparin
5 mm^3 bone marrow core biopsy, fresh tissue or frozen tissue in MPLN RPMI
3.0 mL (min. 2.0 mL) FNA in MPLN RPMI medium
Formalin fixed paraffin embedded tissue

Specimen Stability
EDTA whole blood or bone marrow stable at 18-25°C for 72 hours
Bone marrow biopsy, fresh tissue or FNA in MPLN RPMI at 2-8°C stable for 72 hours
Frozen tissue at 20°C stable indefinitely
Paraffin embedded tissue at 18-25°C stable indefinitely
Storage & Handling

EDTA whole blood, bone marrow, bone marrow biopsy, fresh tissue or FNA, ship in a Styrofoam container with a cool/refrigerated pack (Do not allow the cool pack to directly contact the sample.)
Frozen tissue, ship on dry ice
Paraffin embedded tissue, ship ambient

Causes for Rejection

Improper specimen labeling; Insufficient sample volume; Clotted specimen; Specimen older than 7 days

Reference Range

No rearrangement detected (germline)

Description

Evaluation for suspected clonal B-cell lymphoproliferations when morphologic, immunochemical, and/or flow cytometry analyses are inconclusive. Molecular IgK testing is a useful complement to IgH analysis for confirmation of clonality in post-germinal center (mature) B-cell neoplasms where clonotypic IgH signatures may not be detected due to somatic hypermutation of VH genes.

References
  1. Berget, E., Helgeland, L., Molven, A., & Vintermyr, O. K. (2011). Detection of clonality in follicular lymphoma using formalin-fixed, paraffin-embedded tissue samples and BIOMED-2 immunoglobulin primers. Journal of Clinical Pathology, 64(1), 37-41. doi:10.1136/jcp.2010.081109.
  2. Invivoscribe. (2017). LymphoTrack IGK Assay -MiSeq. 280355 Rev. F.
  3. Lefranc, M. (2001). Nomenclature of the human immunoglobulin kappa (IGK) genes. Exp Clin Immunogenet., 18(3), 161-174.
  4. Mannu, C., Gazzola, A., Bacci, F., Sabattini, E., Sagramoso, C., Roncolato, F., . . . Artioli. (2011). Use of IGK gene rearrangement analysis for clonality assessment of lymphoid malignancies: a single center experience. American Journal of Blood Research, 1(2), 167-174. Retrieved from http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3301430/
  5. Payne, K., Wright, P., Grant, J. W., Huang, Y., Hamoudi, R., Bacon, C. M., . . . Liu, H. (2011). BIOMED-2 PCR assays for IGK gene rearrangements are essential for B-cell clonality analysis in follicular lymphoma. British Journal of Haematology, 155(1), 84-92. doi:10.1111/j.1365-2141.2011.08803.x
  6. Sah, S., Chen, L., Houghton, J., Kemppainen, J., Marko, A. C., Zeigler, R., & Latham, G. J. (2013). Functional DNA quantification guides accurate next-generation sequencing mutation detection in formalin-fixed, paraffin-embedded tumor biopsies. Genome Medicine, 5(8), 77.
  7. van Dongen, J., Langerak, A., Brüggemann, M., Evans, P., Hummel, M., Lavender, F., . . . Macintyre, E. (2003). Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: Report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia, 17, 2257-2317.
  8. van Krieken, J., Langerak, A., Macintyre, E., Kneba, M., Hodges, E., García-Sanz, R., . . . Van Dongen, J. (2007). Improved reliability of lymphoma diagnostics via PCR-based clonality testing: Report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia, 21, 201-206.