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B-cell IGK Gene Rearrangement by Next Generation Sequencing

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

10 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
3 slides (3-5 uM) per marker on adhesion glass

Specimen Stability
EDTA whole blood or bone marrow stable at room temperature 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 stable indefinitely at room temperature
Storage & Handling

Frozen tissue, ship on dry ice
All other specimen types, ship ambient. Protect from extreme temperature with an ice pack. Separate ice pack from specimen.

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.