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B-cell IGH Gene Rearrangement by NGS

Test Code

M BCELL

Test Synonyms

B-cell gene rearrangement

Associations

B-cell leukemia, Lymphoma

Methodology

Next Generation Sequencing (NGS)

Turnaround Time

Assay performed weekly

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 frozen 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

Immunoglobulin Heavy Chain Gene (IGH) V-J Regions

Description

Evaluation for suspected clonal B-cell lymphoproliferations when morphologic, immunochemical and/or flow cytometry analyses are inconclusive.

References
  1. Berget, E., Molven, A., Løkeland, T., Helgeland, L., & Vintermyr, O. K. (2015). IGHV gene usage and mutational status in follicular lymphoma: Correlations with prognosis and patient age. Leukemia Research, 7(39), 702-708.
  2. Invivoscribe. (2017). LymphoTrack IGH (FR1,FR2,&FR3) Assays -MiSeq. 280377 Rev. E.
  3. 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/
  4. Meier, V. S., Rufle, A., & Gudat, F. (2001). Simultaneous Evaluation of T- and B-Cell Clonality, t(11;14) and t(14;18), in a Single Reaction by a Four-Color Multiplex Polymerase Chain Reaction Assay and Automated High-Resolution Fragment Analysis. The American Journal of Pathology, 159(6), 2031-2043. doi:10.1016/S0002-9440(10)63055-6
  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.