Alberta Precision Laboratories

--- 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:

BCR/ABL1 t(9;22) Translocation (Synonym: BCR-ABL1, Qualitative BCR/ABL1, t(9;22) BCR/ABL1 Initial, Philadelphia Chromosome Initial, Ph1 Chromosome by Nested PCR, Quantitative BCR/ABL1, Q-PCR, QPCRPH1, Quantitative PCR, BCR/ABL1 Transcripts by Quantitative PCR)

Clinical Indications:

Qualitative testing is performed for the investigation of myeloproliferative neoplasms and/or acute leukemias.
Quantitative testing is performed for treatment monitoring in CML and acute leukemias.

Test Includes:

Gene: BCR/ABL1

Ordering Alert:

BCR-ABL1 Kinase Domain Mutation is not the same test as BCR-ABL1 Transcripts by Quantitative PCR.

Note: Order BCR/ABL1 t(9;22) Translocation – CHOOSE TESTS if qualitative and/or quantitative testing are/is requested.Order as indicated on the requisition. Do not order both QPCRPH1 and PHLR PB/BM. If both are indicated, order QPCRPH1 only for blood or PHLR BM only for bone marrow.

Collection Alert:

Test cannot be collected by Mobile Collection Service.
Unopened dedicated tubes are required for Molecular testing.

LABID (Connect Care):

LAB4373

Specimen Type:

Bone marrow, Peripheral blood

Specimen Source:

Indicate a specimen source.

Primary Container:

See Specimen Collection Requirements

Minimum Collection Volume:

See Specimen Collection Requirements

Specimen Collection Requirements:

Molecular Pathology North	Molecular Pathology South DSC
Bone Marrow Collect one EDTA (lavender top) tube containing a minimum of 1.0 mL of bone marrow and Collect one PAXgene^® Bone Marrow RNA tube containing a maximum of 2.0 mL of bone marrow. Ensure to follow the specific specimen collection requirements: Place 1.0 to 2.0 mL of bone marrow into the EDTA tube and not more than 2.0 mL of bone marrow into the PAXgene® tube. Order of draw is not important.	Bone Marrow Collect one EDTA (lavender top) tube containing a minimum of 1.0 mL bone marrow.
Peripheral Blood Collect two EDTA tubes each containing a minimum of 1.0 mL of blood and Collect one PAXgene^® Blood RNA tube containing a minimum of 2.5 mL of blood. Ensure to follow the specific specimen collection requirements: Using a butterfly needle and holding the tubes below the arm, first collect 2 EDTA tubes, then the PAXgene® Blood RNA tube. Collection must be in the correct order of draw to ensure accurate white count determination.	Peripheral Blood Collect two EDTA tubes each containing a minimum of 4.0 mL of blood. Do not collect on Friday, Saturday or Sunday. Do not collect on the day preceding a statutory holiday. Read transportation requirements before collection.

Test Resources:

Submit request using the applicable requisition or Connect Care online order entry:

Molecular Pathology North		Molecular Pathology South DSC
	Molecular Pathology Requisition
			Calgary Bone Marrow Collections: Bone Marrow Pathology Requisition
			Calgary Peripheral Blood Collections: Molecular Hematology Requisition

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

Stability and Storage:

Refrigerated until ready for transport.

Processing:

Molecular Pathology North

Molecular Pathology South DSC

Immediately following collection, place all tubes ON ICE.
Do not centrifuge, aliquot or freeze.

For Peripheral Blood:

Send one EDTA tube for a CBC to be ordered and processed as per local handling practices.
Send one EDTA and the PAXgene^® Blood RNA tube to Molecular Pathology.

For Peripheral blood only:

Immediately following collection, place all tubes ON ICE or refrigerate.

For Peripheral blood and Bone Marrow:

Do not centrifuge, aliquot or freeze.

Transportation:

Molecular Pathology North

Molecular Pathology South DSC

Transport specimens as soon as possible, on ice or cold pack.
Molecular Pathology must receive the specimen within 24 hours of collection.
If transport is anticipated to be greater than 24 hours after collection, contact Molecular Pathology.

For Peripheral Blood:

Transport specimens as soon as possible on ice or cold pack.

For Peripheral blood and Bone Marrow:

Specimen must be received in Molecular Pathology South DSC by 15:00 the day following collection.
Specimen will not be accepted if it arrives on Saturday, Sunday or on a statutory holiday
Test may be cancelled if specimen is received by Molecular Hematology more than 48 hours after collection.

Method and Interpretation of Results:

Molecular Pathology North
Qualitative test method: PCR followed by capillary electrophoresis

The sample is reversed transcribed and the resultant cDNA is subjected to PCR. This qualitative, lab-developed assay is designed as a multiplex PCR. The amplified DNA is then run on the 3500xL genetic analyzer for detection and analysis.
The assay is capable of detecting nine different BCR/ABL1 fusion transcripts: b2a2, b3a2, e1a2 (most commonly found in CML and ALL) as well as b2a3, b3a3, e6a2, e8a2, e1a3 and e19a2.
BCR amplification is also measured in every sample to test whether a sample has sufficient quantity and quality of RNA.
The limit of detection (analytical sensitivity) for b2a2, b3a2 and e1a2 fusions was determined to be 3.5 log reduction.

Quantitative p210 BCR/ABL1 test method: Real-time PCR

cDNA synthesis and real-time quantitation of BCR-ABL1 gene transcripts was performed using the BCR-ABL Mbcr IS-MMR Kit purchased from QIAGEN (Hilden, Germany). Appropriate positive and negative controls were used.
Results are reported on the International Scale (IS) which is calibrated against the first NIBSC WHO IS standard (ref. 09/138).
The analytical sensitivity of the UAH Molecular Pathology assay is 0.0032% (4.5 Log Reduction).

Molecular Pathology South DSC
Qualitative test method: Nested PCR followed by TapeStation^® electrophoresis

The most common BCR-ABL1 RNA transcripts associated with Philadelphia chromosome positive leukemia (CML/ALL) are detected by reverse transcription directed, nested PCR analysis.
Primary PCR reactions have a detection limit of approximately 1 abnormal cell in a background of 500 normal cells (major breakpoint – BCR-e14/ABL1 and BCR-e13/ABL1) and 1 abnormal cell in a background of 10 normal cells (minor breakpoint – BCR-e1/ABL1) while nested, secondary PCR reactions have a detection limit of approximately 1 abnormal cell in a background of 1x106 normal cells (major breakpoint) and 1 abnormal cell in a background of 1x105 normal cells (minor breakpoint).
This assay does not detect BCR-ABL1 positive cells with rare translocations outside the major and minor breakpoint regions assessed in these PCR assays.

Quantitative p210 BCR/ABL1 test method: test method: Real-time PCR

cDNA synthesis and quantitative PCR was performed using a IS-MMR kit from Ipsogen Inc. detecting transcripts from the t(9;22) major breakpoint, utilizing TaqMan chemistry (TaKaRa) on the ABI 7500 real time PCR machine.
The results are expressed as a percent normalized copy number (%NCN) ratio of BCR-ABL1/ ABL1, calibrated using assigned value IS-MMR reference material derived from the first NIBSC WHO primary reference standard on the International Scale (IS-MMR 3.0= 0.1%). Treatment response over time is expressed as log reduction from 100% NCN ratio at diagnosis (IS).
Routine test sensitivity is 3 x 10E-3 % (IS-MMR=4.5).

Quantitative p190 BCR/ABL1 test method: test method: Real-time PCR

cDNA synthesis and quantitative PCR was performed using a kit from Ipsogen Inc. detecting transcripts from the t(9;22) minor breakpoint, utilizing Taq Man chemistry (ABI) on the ABI 7500 real time PCR machine.
The results are expressed as a ratio of BCR-ABL1/ ABL1. Because m-bcr breakpoint is rare in CML patients, treatment response over time is expressed as log reduction in the average BCR-ABL1/ ABL1 ratio at ALL diagnosis (0.68, Europe Against Cancer Program).
Test sensitivity is routinely 2x10-4.

Routine Turn Around Time:

Molecular Pathology North	Molecular Pathology South DSC
Qualitative Acute myeloid and lymphocytic leukemias: 3 business days (5 calendar days) Other samples: 8 business days (10 calendar days)	Qualitative New Adult lymphocytic leukemias: 5 calendar days Other samples: 4 weeks
Quantitative 3 weeks	Quantitative IS-MMR calibrated BCR-ABL1: 3 weeks Minor breakpoint BCR-ABL1: 4 weeks

Testing Schedule:

Batched

Testing Area:

Molecular Pathology

Performing Site:

University of Alberta Hospital or Diagnostic and Scientific Center

Additional Test Information:

PAXgene® collection tubes are available upon request.
Submit an order as per online Client Order Form Submission process.
Details in following document: RR01-005 Ordering Supplies from Reagent Room

Contact Comments:

Molecular Pathology North	Molecular Pathology South DSC
780-407-6648	403-770-3699

Last Updated On:

Friday, March 1, 2024

Date of Last Review:

Feb 23 2024 12:00AM

Registered Trademark Comments:

PAXgene^® is a trademark of PreAnalytiX GmbH.
TapeStation^® is a registered trademark of Agilent Technologies, Inc.