--- Query Results --- | |||||
| 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: | IgH Somatic Hypermutation Status (Synonyms: IGH-SH, IGHV, B Cell Receptor Gene Sequencing, Immunoglobulin Heavy Chain Sequencing) | ||||
| Clinical Indications: | For patients with chronic lymphocytic leukemia (CLL) and small lymphocytic lymphoma (SLL) to determine somatic mutation status of the variable gene sequence in the IgH gene. | ||||
| Collection Alert: | Unopened dedicated tubes are required for Molecular Pathology testing. | ||||
| LABID (Connect Care): | LAB4366 | ||||
| Test Code: |
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| Specimen Type: | Blood | ||||
| Primary Container: | Lavender top EDTA | ||||
| Alternate Containers: | Yellow top ACD tube | ||||
| Minimum Collection Volume: | 1.0 mL whole blood | ||||
| Specimen Collection Requirements: | Collect one EDTA (preferred) or one ACD tube. | ||||
| Test Resources: | Submit requests using requisition or Connect Care online order entry:
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| Stability and Storage: | Refrigerate: for up to 3 days | ||||
| Processing: | Do not centrifuge or aliquot. Do not freeze. Keep blood specimens refrigerated until ready to transport. | ||||
| Transportation: | Transport at room temperature to Pathology Department at Foothills Medical Center. | ||||
| Method: | PCR followed by gel isolation and Sanger Sequencing | ||||
| Method and Interpretation of Results: | DNA is extracted using the QIAcube® extractor from Qiagen (Valencia, CA.). The integrity of the DNA is assessed using a Specimen Control Size Ladder detecting four different fragment sizes. Appropriate polyclonal and monoclonal controls are used. A no template control is also included. Samples are run in duplicate. 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 agarose gel electrophoresis and gel isolation of the clonal PCR products. Sanger sequencing is performed on the gel isolated PCR products and the output sequenced is submitted to an online database to determine the mutational status. This assay does not detect 100% clonal cell populations and cannot detect less than 5 positive cells/100 normal cells. Samples must pass assessment with the Specimen Control Size Ladder to be considered suitable for IgH Somatic Hypermutation analysis. | ||||
| Routine Turn Around Time: | 10 days | ||||
| Testing Schedule: | Weekly | ||||
| Reference Interval: | The presence of IGH somatic hypermutation (SHM) is defined as greater or equal to 2% difference from the germline variable (V) gene sequence, whereas less than 2% difference is considered evidence of no somatic hypermutation. | ||||
| Testing Area: | Molecular Pathology | ||||
| Performing Site: | Foothills Medical Center | ||||
| Contact Comments: | Molecular Pathology South 403-220-4240 | ||||
| Last Updated On: | Monday, July 27, 2020 | ||||
| Date of Last Review: | Aug 1 2022 12:00AM | ||||
| Registered Trademark Comments: | QIAcube® is a registered trademark of the QIAGEN Group. | ||||
