Immunoglobulin Heavy Chain (IGH) Clonality Detection by PCR and Capillary Electrophoresis
IGH Gene Clonality Detection, B-cell Receptor IGH Gene Rearrangement
Molecular Diagnostics Laboratory
Clinical Indication and Relevance
The assay is useful to assist with the diagnosis of B-cell malignancies. The assay can also be used to detect minimal residual disease in follow-up samples.
DNA is isolated and amplified by PCR using BIOMED-2 primers targeting the VH framework 1, 2, 3, DH and JH sequences of the IGH gene. The gene rearrangements are detected by analyzing the PCR products by capillary gel electrophoresis. The assay sensitivity for detection of clonal B-cell populations is 5% of lymphocytes.
Minimum Specimen Requirements
- Peripheral blood (PB): 3-5 mL, in purple top (sodium EDTA) tube.
- Bone marrow (BM): 1-3 mL, drawn into a syringe containing anticoagulant and then
delivered in purple top tube (EDTA).
- Fresh or frozen tissue: fresh tissue should be obtained in a sterile manner, and a minimum
of tissue is required. Fresh tissue should be placed in culture medium for transport
or snap frozen (see transport section below).
- Formalin-fixed, paraffin-embedded (FFPE) tissue blocks: send tissue block to the lab, or
contact lab for instructions regarding cutting paraffin sections for molecular studies.
- PB or BM sample: Deliver immediately at 2-8°C (wet ice or cold packs). Do not freeze.
- Fresh tissue samples should be delivered at room temperature in RPMI culture medium within 3 hours of collection, or snap frozen in liquid nitrogen at -70°C and packed in
dry ice for delivery. Please do not allow frozen tissues to thaw.
- Formalin-fixed paraffin embedded (FFPE) tissue blocks and slides can be delivered at controlled temperatures, not to exceed 30C.
PB or BM samples: ambient – 1 hour; refrigerated – 48 hours.
- Serum or plasma; frozen PB or BM; clotted blood; severely hemolyzed samples
- Paraffin tissue fixed in Zenker’s, B5, or Bouin’s fixatives
- Bone marrow biopsy decalcified in formic acid
Five to seven working days.View Requisition Sheet