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HelpTest Code CXLPL CXCR4 Mutation Analysis, Somatic, Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, Varies
Ordering Guidance
It is strongly recommended that this test be used in the context of results of MYD88 / MYD88, L265P, Somatic Gene Mutation, DNA Allele-Specific PCR, Varies. If MYD88 has not been previously performed, consider LPLFX / Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, MYD88 L265P with Reflex to CXCR4, Varies during evaluation of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia.
Shipping Instructions
Whole blood or bone marrow specimens must arrive within 10 days of collection.
Necessary Information
The following information is required:
1. Pertinent clinical history
2. Clinical or morphologic suspicion
3. Date and time of collection
4. Specimen source
Specimen Required
Submit only 1 of the following specimens:
Preferred
Specimen Type: Bone marrow aspirate
Container/Tube:
Preferred: Lavender top (EDTA)
Acceptable: Yellow top (ACD), green top (sodium heparin)
Specimen Volume: 2 mL
Collection Instructions:
1. Invert several times to mix bone marrow.
2. Send bone marrow specimen in original tube. Do not aliquot.
3. Label specimen as bone marrow.
Specimen Stability Information: Ambient (preferred) 10 days /Refrigerated 10 days
Specimen Type: Whole blood
Container/Tube:
Preferred: Lavender top (EDTA)
Acceptable: Yellow top (ACD), green top (sodium heparin)
Specimen Volume: 3 mL
Collection Instructions:
1. Invert several times to mix blood.
2. Send whole blood specimen in original tube. Do not aliquot.
3. Label specimen as blood.
Specimen Stability Information: Ambient (preferred) 10 days/Refrigerated 10 days
Specimen Type: Paraffin-embedded tissue
Container/ Tube: Paraffin block
Collection Instructions:
1. Decalcified specimens (eg, bone marrow core biopsies) are not acceptable.
2. Indicate specimen source.
Specimen Stability Information: Ambient
Additional Information: If the quality of the biopsy specimen is poor, testing should not be ordered. Testing may be canceled if DNA requirements are inadequate.
Acceptable
Specimen Type: Tissue slide
Slides: 20 Unstained slides
Container/ Tube: Transport in plastic slide holders
Collection Instructions:
1. Send 20 unstained, nonbaked slides with 5-micron thick sections of tissue.
2. Decalcified specimens (eg, bone marrow core biopsies) are not acceptable.
3. Indicate specimen source.
Specimen Stability Information: Ambient
Additional Information: Testing may be canceled if resultant extracted DNA does not meet concentration requirements.
Specimen Type: Frozen tissue
Container/Tube: Plastic container
Specimen Volume: 100 mg
Collection Instructions:
1. Freeze tissue within 1 hour of collection
2. Indicate specimen source.
Specimen Stability Information: Frozen
Additional Information: Testing may be canceled if resultant extracted DNA does not meet concentration requirements.
Specimen Type: Extracted DNA
Container/Tube: 1.5- to 2-mL tube
Specimen Volume: Entire specimen
Collection Instructions:
1. DNA must be extracted within 7 days of collection.
2. Label specimen as extracted DNA and source of specimen.
3. Provide volume and concentration of DNA on label.
Specimen Stability Information: Frozen (preferred)/Refrigerated/Ambient
Additional Information: DNA must be extracted in a CLIA-certified laboratory or equivalent and must be extracted from a specimen type listed as acceptable for this test (including applicable anticoagulants). We cannot guarantee that all extraction methods are compatible with this test. If testing fails, one repeat will be attempted, and if unsuccessful, the test will be reported as failed and a charge will be applied.
Forms
1. Hematopathology Patient Information (T676)
2. If not ordering electronically, complete, print, and send a Hematopathology/Cytogenetics Test Request (T726) with the specimen.
Useful For
Aiding in the prognosis and clinical management of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia
Genetics Test Information
This test detects gene mutations within the C-terminal end of the CXCR4 gene that are commonly found in association with MYD88 L265P mutations in cases of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia.
Special Instructions
Method Name
Bridged Nucleic Acids (BNA) Clamp Sanger Sequencing Technology/Routine Sanger Sequencing (BNAClamp is utilized pursuant to a license agreement with BNA Inc)
Reporting Name
CXCR4 Mutation in B-cell LymphomaSpecimen Type
VariesSpecimen Minimum Volume
Whole blood, bone marrow aspirate, 0.5 mL; Frozen tissue: 50 mg; Extracted DNA: 50 mcL at 20 ng/mcL; Tissue slides: 10 unstained slides
Specimen Stability Information
| Specimen Type | Temperature | Time |
|---|---|---|
| Varies | Varies | 10 days |
Reject Due To
| Gross hemolysis | Reject |
| B5-fixed tissues | Reject |
| Decalcified bone marrow core biopsies | Reject |
| Frozen tissue | Reject |
| Methanol acetic acid (MAA)-fixed pellets | Reject |
| Moderately to severely clotted | Reject |
| Paraffin shavings | Reject |
Clinical Information
Lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM) is a B-cell lymphoma characterized by an aberrant accumulation of malignant lymphoplasmacytic cells in the bone marrow, lymph nodes, and spleen. It is a B-cell neoplasm that can exhibit excess production of serum IgM symptoms related to hyperviscosity, tissue filtration, and autoimmune-related pathology. CXCR4 mutations are identified in approximately 30% to 40% of patients with LPL/WM and are almost always associated with MYD88 L265P, which is highly prevalent in this neoplasm. The status of CXCR4 mutations in the context of MYD88 L265P is clinically relevant as important determinants of clinical presentation, overall survival, and therapeutic response to ibrutinib. A MYD88-L265P/CXCR4-WHIM (C-terminus nonsense/frameshift mutations) molecular signature is associated with intermediate to high bone marrow disease burden and serum IgM levels, less adenopathy, and intermediate response to ibrutinib in previously treated patients. A MYD88-L265P/CXCR4-WT (wildtype) molecular signature is associated with intermediate bone marrow disease burden and serum IgM levels, more adenopathy, and highest response to ibrutinib in previously treated patients. A MYD88-WT/CXCR4-WT molecular signature is associated with inferior overall survival, lower response to ibrutinib therapy in previously treated patients, and lower bone marrow disease burden in comparison to those harboring a MYD88-L265 mutation.
Reference Values
Mutations present or absent in the test region c. 898-1059 (amino acids 300-353) of the CXCR4 gene (NCBI NM_003467.2, GRCh37)
Interpretation
Mutation present or not detected; an interpretive report will be issued.
Cautions
This test is a targeted assay for the C-terminal end of the CXCR4 gene only. It examines c.898-1059 of the CXCR4 gene (NCBI NM_003467.2 GRCh37) and does not detect mutations outside this region. A 1% analytical sensitivity was established at 50 ng DNA input for the hotspot mutations c.1013C>G/A only, which uses bridged nucleic acids-clamped Sanger sequencing, and DNA not meeting established criteria can lead to false-negative results. In the extremely rare event that a rare polymorphism, insertion, or deletion occurs at the Sanger sequencing primer binding sites, in cis with c.1013C>G/A, data can yield a failed result. Routine Sanger sequencing is used to interrogate other mutations in the tested region with a 15% to 20% analytical sensitivity. The analytical sensitivity of the assay can be affected by a variety of factors, including biologic availability (ie, tumor burden), fixation of paraffin-embedded specimens, rare polymorphisms, insertions, or deletions at the primer binding sites, or nonspecific polymerase chain reaction interferences.
Method Description
The C-terminal end of CXCR4 (NM_003467.2, c.898-1059) is amplified from extracted genomic DNA by polymerase chain reaction, followed by Sanger sequencing and capillary electrophoresis analysis. Review of the sequence data is performed using a combination of automated calls and manual inspection.(Unpublished Mayo method)
The hotspot mutations c.1013C>G/A (p.S338X) are examined using bridged nucleic acids clamped Sanger sequencing with an analytic sensitivity of 1%. All other genetic mutations in the test region are examined by routine Sanger sequencing with an analytic sensitivity of 15% to 20%.(Unpublished Mayo method)
Day(s) Performed
Monday through Friday
Report Available
7 to 10 daysSpecimen Retention Time
Bone marrow aspirate/Whole blood/Fresh/Frozen Tissue: 2 weeks; Extracted DNA: 3 months; FFPE tissue: Unused portions of blocks will be returned to the client. Unstained slides: Not retainedPerforming Laboratory
Mayo Clinic Laboratories in Rochester
Test Classification
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.CPT Code Information
81479
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| CXLPL | CXCR4 Mutation in B-cell Lymphoma | In Process |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| MP032 | Specimen Type | 31208-2 |
| 113436 | CXLPL Result | 59465-5 |
| 38287 | Final Diagnosis | 50398-7 |