Alberta Precision Laboratories

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Organization:

Alberta Precision Laboratories

Message Alert:

PLEASE NOTE: This Test Directory is in an interim state and due to changes pertaining to the DynaLIFE transition, performing site for routing has not been updated. Please refer to Laboratory Information System for appropriate routing.

Test Name/Synonym:

B-cell Clonality (IgH Gene Rearrangement) (Synonyms: B Cell Gene Rearrangement, B Cell Clonality, Immunoglobulin Heavy Chain Gene Rearrangement)

Clinical Indications:

For the investigation of lymphoproliferative disorders.

Collection Alert:

Unopened dedicated tubes are required for Molecular Pathology testing.

LABID (Connect Care):

LAB4195

Specimen Type:

Blood, Bone Marrow, Paraffin-Embedded Tissue, Fresh tissue

Primary Container:

See Specimen Collection Requirements

Specimen Collection Requirements:

Molecular Pathology North	Molecular Pathology South
Peripheral Blood Collect two lavender top EDTA tubes, each containing 1.0 mL blood.	Peripheral Blood Collect one lavender top EDTA tube (preferred) or one pale yellow top ACD tube, containing 1.0 mL blood.
Bone Marrow Collect one EDTA tube containing 2.0 mL bone marrow.	Bone Marrow Bone Marrow from Flow Cytometry supplied in Bone Marrow Media (BMM).
Paraffin Embedded Tissue A formalin-fixed, paraffin-embedded tissue block is preferred. Alternatively, one slide stained with Hematoxylin & Eosin and five 10-micron non-baked, unstained slides can be submitted. Please provide a copy of the corresponding pathology report.	Paraffin Embedded Tissue Can be submitted as whole block, precut scrolls, or as blank unstained sections on coated slides. Representative H&E to accompany the specimen.
	Tumor/Tissue Submit 5.0 mg tissue in flow media.

Test Resources:

Submit request using the applicable requisition:

Molecular Pathology North		Molecular Pathology South
	Molecular Pathology Requisition		Molecular Pathology Requisition

Please ensure all required information, including patient's clinical history/indication, is provided on the requisition.

Stability and Storage:

Refrigerated for up to 3 days: blood, bone marrow and fresh tissue in flow media specimens
Room temperature: paraffin embedded tissue

Processing:

Molecular Pathology North	Molecular Pathology South
Do not centrifuge, aliquot or freeze. Keep blood and bone marrow specimens refrigerated until ready to transport. For Peripheral Blood: Submit one EDTA tube for a CBC to be ordered and processed as per local handling practices. Submit one EDTA to Molecular Pathology.	Do not centrifuge, aliquot or freeze. Keep blood, bone marrow and fresh tissue samples (in media) refrigerated until ready to transport.

Transportation:

Molecular Pathology North

Molecular Pathology South

Blood and Bone Marrow

Must be received in Molecular Pathology within 3 days of collection.
Transport at room temperature to Molecular Pathology via University of Alberta Hospital Specimen Control WMC 4B2.10.

Paraffin Embedded Tissue

Transport at room temperature to Molecular Pathology at University of Alberta Hospital

Transport specimens at ambient (room) temperature to Pathology Department at Foothills Medical Center.

Method:

PCR followed by capillary electrophoresis

Method and Interpretation of Results:

Molecular Pathology North
PCR is performed using Invitrogen Platinum High Fidelity Taq with fluorescence-labelled primers in three different mixes. The PCR products are subjected to capillary electrophoresis on an Applied Biosystems 3130/3500XL Genetic Analyzer using fragment analysis GeneMapper software. The integrity of the DNA is assessed using a Specimen Control Size Ladder detecting four different fragment sizes. Appropriate polyclonal and monoclonal controls (B-cell lymphoma cell lines) are used. A no template control is also included.

This assay does not detect 100% clonal cell populations and cannot detect less than 5 positive cells/95 normal cells.

The clinical sensitivity of this assay is determined to be 75% and clinical specificity is 100%.

Molecular Pathology South
A commercially available kit from InVivoscribe Technologies (San Diego, California) is used that targets rearrangements in IgH for the identification of clonal B-cell populations. PCR is performed using Amplitaq Gold DNA polymerase with appropriate fluorescence-labeled primers in five different mixes. The PCR products are subjected to capillary electrophoresis on an Applied Biosystems 3500 Genetic Analyzer using fragment analysis GeneMapper software.

This assay does not detect 100% clonal cell populations and cannot detect less than 1 positive cells/100 normal cells.

Samples must pass assessment with the Specimen Control Size Ladder to be considered suitable for IgH gene rearrangement analysis.

Routine Turn Around Time:

Molecular Pathology North	Molecular Pathology South
2 weeks	10 days

Testing Schedule:

Twice per week (batched)

Testing Area:

Molecular Pathology

Performing Site:

University of Alberta Hospital or Foothills Medical Centre

Contact Comments:

Molecular Pathology North	Molecular Pathology South
780-407-6648	403-220-4240

Last Updated On:

Thursday, December 5, 2024

Date of Last Review:

Feb 22 2024 12:00AM