--- Query Results ---
Program:	Genetics and Genomics
Test Name:	T Cell Receptor Gene Rearrangement
Alternate Test Name:	TCR Gene Rearrangement
Performing Site:	University of Alberta Hospital
Performing Dept:	Molecular Pathology
Availability:	Test set-up on Monday and Thursday (excluding statutory holidays).
TAT:	TCR gamma 7 days, TCR beta 14 days
Preferred Tube/Container:	See Specimen Requirements
Specimen Requirements:	For Blood Samples: Unopened dedicated tubes are required for Molecular Pathology testing. 1. Any peripheral blood requests will require a CBC and differential to be ordered and processed as per local handling practices. 2. Collect 2 x 4 mL EDTA (lavender) tubes. One tube for CBC/differential and one tube for Molecular Pathology. For Bone Marrow Samples: Unopened dedicated tubes are required for Molecular Pathology testing. 1. Collect 1 x 4 mL EDTA (lavender) tube. 2. Minimum 2 mL of bone marrow required. For Paraffin-Embedded Tissue: 1. A formalin-fixed, paraffin-embedded tissue block is preferred. 2. Alternatively, one slide stained with Hematoxylin & Eosin and five 10-micron non-baked, unstained slides can be submitted. 3. Please provide a copy of the corresponding pathology report.
Specimen Processing:	Do not spin. Do not freeze. Keep samples refrigerated until ready to transport.
Specimen Handling:	Whole blood samples must be received in Molecular Pathology within 3 days of collection. Transport samples at ambient temperature.
Requisition/Form:	All requests MUST be submitted on a Molecular Pathology Requisition. Please provide any pertinent clinical history on the requisition.
Order Restrictions:	Test requests are restricted to hematopathologists and lymphoma pathologists.
Indications:	For the investigation of lymphoproliferative disorders.
Gene:	TCR
Method:	PCR followed by capillary electrophoresis.
Method Details:	DNA is extracted using the QIAsymphony DSP DNA Mini Kit 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 (T-cell lymphoma cell line) controls are used. A no template control is also included. These assays have been validated by a parallel study with another institution. T-Cell Receptor Gamma Assay A four-color, multiplex, PCR assay is performed using Invitrogen Platinum High Fidelity Taq with appropriate fluorescence-labeled primers detecting four different fragment sizes. The PCR products are subjected to capillary electrophoresis on an Applied Biosystems 3130/3500XL Genetic Analyzer using fragment analysis GeneMapper software. Previously published primers for the four-color TCR gamma PCR Assay (Vega et al, Am J Clin Pathol 2001;116:17-24) are utilized in this method. Due to the sensitivity of this assay, pseudoclonality can be seen occasionally in some inflammatory conditions. These clones are usually not reproducible when a repeat sample is obtained. This assay cannot detect less than 5 positive cells/95 normal cells. The clinical sensitivity of this assay is determined to be 87.5% and clinical specificity is 95% in our laboratory. T-Cell Receptor Beta Assay A commercially available kit from InVivoscribe Technologies (San Diego, California) is used. PCR is performed using Invitrogen Platinum High Fidelity Taq with appropriate fluorescence-labeled 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. 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 52% and clinical specificity is 84% in our laboratory.
Last Updated On:	Wednesday, November 14, 2018
Date of Last Review:	Jan 2 2018 12:00AM