Test Catalog

Test Id : LPLFX

Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, MYD88 L265P with Reflex to CXCR4, Varies

Useful For
Suggests clinical disorders or settings where the test may be helpful

Establishing the diagnosis of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM)

 

Helping distinguish LPL/WM low-grade B-cell lymphoma from other subtypes

 

Aiding in the prognostication and clinical management of LPL/WM

Highlights

This test offers highly sensitive detection of the well-characterized hotspot variants c.1013C->G/A, p.S338X and routine Sanger sequencing for other variants in the C-terminus region.

Reflex Tests
Lists tests that may or may not be performed, at an additional charge, depending on the result and interpretation of the initial tests.

Test Id Reporting Name Available Separately Always Performed
CXCFX CXCR4, Gene Mutation, Reflex Yes, (order CXLPL), Bill Only No

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

The algorithm starts with the sensitive MYD88 L265P testing by allele-specific polymerase chain reaction. If a MYD88 L265P variant is detected, additional CXCR4 testing will be performed. If a MYD88 L265P variant is not detected, the algorithm ends and no further testing is necessary.

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Name
A short description of the method used to perform the test

Allele-Specific Polymerase Chain Reaction (AS-PCR), Bridged Nucleic Acids (BNA) Clamp Sanger Sequencing and Routine Sanger Sequencing

(BNAClamp is utilized pursuant to a license agreement with BNA Inc.)

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

Reflex Testing of MYD88 and CXCR4

Aliases
Lists additional common names for a test, as an aid in searching

B-cell lymphoma

L265P

LPL/WM

Lymphoplasmacytic lymphoma

MYD88

CXCR4

Waldenstrom Macroglobulinemia

CXLPL

S338X

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

The algorithm starts with the sensitive MYD88 L265P testing by allele-specific polymerase chain reaction. If a MYD88 L265P variant is detected, additional CXCR4 testing will be performed. If a MYD88 L265P variant is not detected, the algorithm ends and no further testing is necessary.

Specimen Type
Describes the specimen type validated for testing

Varies

Shipping Instructions

Peripheral 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

ORDER QUESTIONS AND ANSWERS

Question ID Description Answers
MP042 Specimen Type

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Submit only 1 of the following specimens:

 

Preferred:

Specimen Type: Bone marrow

Container/Tube:

Preferred: Lavender top (EDTA)

Acceptable: Yellow top (ACD solution B)

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)/Refrigerated

 

Specimen Type: Paraffin-embedded tissue

Container/Tube: Paraffin block

Specimen Stability: Ambient

 

Specimen Type: Paraffin-embedded bone marrow aspirate clot

Container/Tube: Paraffin block

Specimen Stability: Ambient

 

Specimen Type: Tissue

Slides: Unstained slides

Specimen Volume: 10 slides

Additional Information: Tissue must demonstrate involvement by a hematologic neoplasm (eg, acute myelocytic leukemia), not solid tumors.

Specimen Stability Information: Ambient

 

Acceptable:

Specimen Type: Peripheral blood

Container/Tube:

Preferred: Lavender top (EDTA)

Acceptable: Yellow top (ACD solution B)

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)/Refrigerated

 

Specimen Type: Extracted DNA

Container/Tube: 1.5- to 2-mL tube with indication of volume and concentration of the DNA

Specimen Volume: Entire specimen

Collection Instructions: Label specimen as extracted DNA and list the specimen source. Include indication of volume and concentration of the DNA.

Specimen Stability Information: Frozen (preferred)/Refrigerated/Ambient

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Forms

1. Hematopathology Patient Information (T676) in Special Instructions

2. If not ordering electronically, complete, print, and send a Hematopathology/Cytogenetics Test Request (T726) with the specimen.

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

Blood, Bone marrow: 1 mL

Extracted DNA: 20 mcL with a concentration of at least 10 nanograms per mcL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis Reject
B5-fixed tissues
Decalcified bone marrow biopsies
Methanol-acetic acid (MAA)-fixed pellets
Paraffin shavings
Frozen tissue
Moderately to severely clotted
Reject

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Varies Varies 10 days

Useful For
Suggests clinical disorders or settings where the test may be helpful

Establishing the diagnosis of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM)

 

Helping distinguish LPL/WM low-grade B-cell lymphoma from other subtypes

 

Aiding in the prognostication and clinical management of LPL/WM

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

The algorithm starts with the sensitive MYD88 L265P testing by allele-specific polymerase chain reaction. If a MYD88 L265P variant is detected, additional CXCR4 testing will be performed. If a MYD88 L265P variant is not detected, the algorithm ends and no further testing is necessary.

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

The MYD88 L265P abnormality is highly associated (>90%) with the pathologic diagnosis of lymphoplasmacytic lymphoma and the clinical syndrome of Waldenstrom macroglobulinemia (LPL/WM), particularly in the setting of an elevated IgM serum monoclonal paraprotein.

 

CXCR4 variants are identified in approximately 30% to 40% of LPL/WM patients and are almost always in association with MYD88 L265P, which is highly prevalent in this neoplasm. The status of CXCR4 variants 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 variants) 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 (wild type) molecular signature is associated with intermediated 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 variant.

Reference Values
Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.

MYD88 L265P: Variant present or absent based on expected variant PCR product size for the MYD88 gene (NCBI accession NM_002468.4).

 

CXCR4: Variants present or absent in the test region c. 898-1059 (amino acids 300-353) of the CXCR4 gene (NCBI NM_003467.2, GRCh37).

Interpretation
Provides information to assist in interpretation of the test results

Reportable variant present or not detected; an interpretive report will be provided.

Cautions
Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

This MYD88 test is a targeted assay and will not detect any alteration at the MYD88 codon 265 that does not result in the L>P amino acid change. It will also not detect additional MYD88 variants, including insertion or deletion events. The analytical sensitivity of the assay (1% MYD88 L265P in a wild type background) can be affected by a variety of factors, including biologic availability (ie, tumor burden), fixation of paraffin-embedded specimens, or nonspecific polymerase chain reaction (PCR) interferences. Rare cases of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM) have been reported lacking the MYD88 L265P abnormality, so a negative result would not completely exclude this diagnosis but would make the possibility of LPL/WM more unlikely.

 

The reflexed 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 variants outside this region. A 1% analytical sensitivity was established at 50 ng DNA input for the hotspot variants c.1013C>G/A only, which uses bridged nucleic acids (BNA) clamped Sanger sequencing and DNA that does not meet the established criteria can lead to false-negative results. In the extremely rare event that a rare benign variant (ie, polymorphism), insertion, or deletion may occur at the Sanger sequencing primer binding sites, in cis, with a c.1013C>G/A, data can yield a failed result. Routine Sanger sequencing is used to interrogate other variants 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 benign variants, insertions, or deletions at the primer binding sites or nonspecific PCR interferences.

Clinical Reference
Recommendations for in-depth reading of a clinical nature

1. Treon SP, Xu L, Yang G, et al: MYD88 L265P somatic mutation in Waldenstrom's macroglobulinemia. N Engl J Med. 2012 Aug 30;367(9):826-833. doi: 10.1056/NEJMoa1200710

2. Varettoni M, Arcaini L, Zibellini S, et al: Prevalence and clinical significance of the MYD88 (L265P) somatic mutation in Waldenstrom's macroglobulinemia and related lymphoid neoplasms. Blood. 2013 Mar 28;121(13):2522-2528. doi: 10.1182/blood-2012-09-457101

3. Xu L, Hunter ZR, Yang G, et al: MYD88 L265P in Waldenstrom macroglobulinemia, immunoglobulin M monoclonal gammopathy, and other B-cell lymphoproliferative disorders using conventional and quantitative allele-specific polymerase chain reaction. Blood. 2013 Mar 14;121(11):2051-2058. doi: 10.1182/blood-2012-09-454355

4. Poulain S, Roumier C, Decambron A, et al: MYD88 L265P mutation in Waldenstrom macroglobulinemia. Blood. 2013 May 30;121(22):4504-4511. doi: 10.1182/blood-2012-06-436329

5. Gachard N, Parrens M, Soubeyran I, et al: IGHV gene features and MYD88 L265P mutation separate the three marginal zone lymphoma entities and Waldenstrom macroglobulinemia/lymphoplasmacytic lymphomas. Leukemia. 2013 Jan;27(1):183-189. doi: 10.1038/leu.2012.257

6. Ondrejka SL, Lin JJ, Warden DW, et al: MYD88 L265P somatic mutation: its usefulness in the differential diagnosis of bone marrow involvement by B-cell lymphoproliferative disorders. Am J Clin Pathol. 2013 Sept;140(3):387-394. doi: 10.1309/AJCP10ZCLFZGYZIP

7. Hunter Z, Xu L, Yang G, et al: The genomic landscape of Waldenstrom macroglobulinemia is characterized by highly recurring MYD88 and WHIM-like CXCR4 mutations, and small somatic deletions associated with B-cell lymphomagenesis. Blood. 2014 Mar 13;123(11):1637-1646. doi: 10.1182/blood-2013-09-525808

9. Poulain S, Roumier C, Venet-Caillault A, et al: Genomic landscape of CXCR4 mutations in Waldenstrom macroglobulinemia. Clin Cancer Res. 2016 Mar 15;22(6):1480-1488. doi: 10.1158/1078-0432.CCR-15-0646

10. Roccaro A, Sacco A, Jimenez C, et al: C1013G/CXCR4 acts as a driver mutation of tumor progression and modulator of drug resistance in lymphoplasmacytic lymphoma. Blood. 2014 Jun 26;123(26):4120-4131. doi: 10.1182/blood-2014-03-564583

11. Schmidt J, Federmann B, Schindler N, et al: MYD88 L265P and CXCR4 mutations in lymphoplasmacytic lymphoma identify cases with high disease activity. Br J Haematol. 2015 Jun;169(6):795-803. doi: 10.1111/bjh.13361

12. Treon SP, Cao Y, Xu L, et al: Somatic mutations in MYD88 and CXCR4 are determinants of clinical presentation and overall survival in Waldenstrom macroglobulinemia. Blood. 2014 May 1;123(18):2791-2796. doi: 10.1182/blood-2014-01-550905

13. Treon SP, Tripsas CK, Meid K, et al. Ibrutinib in previously treated Waldenstrom's macroglobulinemia. N Engl J Med. 2015 Apr 9;372(15):1430-1440. doi: 10.1056/NEJMoa1501548

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Description
Describes how the test is performed and provides a method-specific reference

Extracted DNA from the clinical specimen is subjected to allele-specific polymerase chain reaction (PCR) using MYD88 exon 5 primers that simultaneously amplify both a wild-type sequence fragment and a fragment containing the specific nucleotide change resulting in L265P, if present. PCR products are visualized by capillary electrophoresis and the presence of mutated and wild type amplicons is determined according to the expected specific PCR product sizes.(Unpublished Mayo method)

 

The C-terminal end of CXCR4 (NM_003467.2, c. 898-1059) is amplified from extracted genomic DNA by PCR, 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 variants c.1013C>G/A (p.S338X) are examined using bridged nucleic acids clamped Sanger sequencing with an analytic sensitivity of 1%. All other genetic variants in the test region are examined by routine Sanger sequencing with an analytic sensitivity of 15% to 20%.(Unpublished Mayo method)

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Friday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

7 to 10 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

DNA: 3 months; Peripheral blood, bone marrow: 2 weeks

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

Test Classification
Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

This test was developed, and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information
Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

81305

LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
LPLFX Reflex Testing of MYD88 and CXCR4 82140-5
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
MP042 Specimen Type 31208-2
601511 LPLFX Reflex Result 82140-5
601510 Final Diagnosis 50398-7

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports