Test Catalog

Test Id : NPM1Q

Nucleophosmin (NPM1) Mutation Analysis, Varies

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

As a prognostic indicator in patients with newly diagnosed acute myelogenous leukemia with normal karyotype and no FLT3 variant and as a leukemia-specific marker of minimal residual disease

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

Both DNA and RNA are extracted. The assay is composed of 2 parts, an RNA-based, sensitive quantitative reverse transcription real-time polymerase chain reaction (RT-PCR) that detects and quantifies the most common altered NPM1 messenger RNA transcripts (A, B, D forms) in acute myeloid leukemia (AML), and a DNA-based qualitative NPM1 exon 12 variant screening by fragment analysis that detects essentially all altered forms reported in AML, including the rare non-A, B, D forms (with lower sensitivity at the DNA level).

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

RNA: Reverse-Transcription Quantitative PCR (RT-qPCR)

DNA: Polymerase Chain Reaction (PCR) with Fragment Analysis by Capillary Gel Electrophoresis

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

NPM1 Mutation Analysis, V

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

NPM1

Nucleophosmin Mutation Analysis

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

Both DNA and RNA are extracted. The assay is composed of 2 parts, an RNA-based, sensitive quantitative reverse transcription real-time polymerase chain reaction (RT-PCR) that detects and quantifies the most common altered NPM1 messenger RNA transcripts (A, B, D forms) in acute myeloid leukemia (AML), and a DNA-based qualitative NPM1 exon 12 variant screening by fragment analysis that detects essentially all altered forms reported in AML, including the rare non-A, B, D forms (with lower sensitivity at the DNA level).

Specimen Type
Describes the specimen type validated for testing

Varies

Shipping Instructions

Refrigerated specimen must arrive within 5 days of collection, and ambient specimens must arrive within 3 days of collection. Collect and package specimen as close to shipping time as possible.

Necessary Information

The following information is required:

1. Pertinent clinical history

2. Clinical or morphologic suspicion

3. Date of collection

4. Specimen source

ORDER QUESTIONS AND ANSWERS

Question ID Description Answers
MP053 Specimen Type Peripheral blood
Bone marrow

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:

 

Specimen Type: Blood

Container/Tube: Lavender top (EDTA) or yellow top (ACD-B)

Specimen Volume: 10 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 Type: Bone marrow

Container/Tube: Lavender top (EDTA) or yellow top (ACD-B)

Specimen Volume: 4 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.

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

Forms

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

Blood: 8 mL

Bone marrow: 2 mL

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

Gross hemolysis Reject
Bone marrow biopsies
Paraffin-embedded bone marrow clots
Slides
Paraffin shavings
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 Refrigerated (preferred) 5 days
Ambient 72 hours

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

As a prognostic indicator in patients with newly diagnosed acute myelogenous leukemia with normal karyotype and no FLT3 variant and as a leukemia-specific marker of minimal residual disease

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

Both DNA and RNA are extracted. The assay is composed of 2 parts, an RNA-based, sensitive quantitative reverse transcription real-time polymerase chain reaction (RT-PCR) that detects and quantifies the most common altered NPM1 messenger RNA transcripts (A, B, D forms) in acute myeloid leukemia (AML), and a DNA-based qualitative NPM1 exon 12 variant screening by fragment analysis that detects essentially all altered forms reported in AML, including the rare non-A, B, D forms (with lower sensitivity at the DNA level).

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

Acute myeloid leukemia (AML) is a genetically heterogeneous group of neoplasms. While cytogenetic aberrations detected at the time of diagnosis are the most used prognostic feature, approximately 50% of AML cases show a normal karyotype, which is considered an intermediate-risk feature. Within this group, FLT3 variants are considered indicators of poor prognosis. However, in the absence of a FLT3 variant, the presence of a NPM1 variant is associated with a more favorable prognosis. A NPM1 alteration is a common finding in de novo AML (25%-30% of cases) and consists of small insertion (typically 4 base pair) or deletion/insertion events involving exon 12. Three variants are highly recurrent, termed types A, B, and D, and together account for approximately 90% of NPM1 alterations in de novo AML. Thus, in patients with newly diagnosed AML, those with normal karyotype, no FLT3 variant, and a NPM1 alteration are considered to have a better prognosis than patients in the same group with neoplasms lacking a NPM1 alteration. Furthermore, the presence of a NPM1 alteration serves as a sensitive marker for evaluating minimal disease and therapeutic response following treatment.

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.

An interpretive report will be provided.

Interpretation
Provides information to assist in interpretation of the test results

The assay incorporates 2 parts: a qualitative screen for exon 12 NPM1 alterations and a quantitative reverse transcription polymerase chain reaction (RT-PCR) to determine the copy number of NPM1 transcripts (relative to ABL1 reference mRNA). This strategy will allow for identification of the NPM1 alteration at diagnosis, as well as a high sensitivity method to monitor patients who are post-therapy for minimal residual disease. Results will therefore be interpreted with integration of the quantitative and qualitative test results in the context of NPM1 alteration type identified at the time of AML diagnosis if available. Because the quantitative RT-PCR component only reliably detects and quantifies the 3 most common variant types (A, B, D), there is a very small possibility that the qualitative assay may indicate the presence of NPM1 alteration, but the quantitative assay will be (falsely) negative. In patients with newly diagnosed acute myeloid leukemia, a normal karyotype, and no FLT3 variant, the presence of NPM1 alteration is an indicator of a more favorable prognosis. Similarly, following chemotherapy, the presence, relative quantity, and trend of change of NPM1 messenger RNA transcript is associated with risk of disease relapse.

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

Because of the design of this assay, a very small number of NPM1 alterations at diagnosis may not be detected by the more targeted quantitative polymerase chain reaction component. In that setting, the qualitative part of the test can be used for limited minimal residual disease assessment, although the sensitivity is much lower (approximately 5% at the DNA level).

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

1. Heath EM, Chan SM, Minden MD, Murphy T, Shlush LI, Schimmer AD: Biological and clinical consequences of NPM1 mutations in AML. Leukemia. 2017 Apr;31(4):798-807. doi: 10.1038/leu.2017.30

2. Kronke J, Schlenck RF, Jensen KO, et al: Monitoring of minimal residual disease in NPM1-mutated acute myeloid leukemia: a study from the German-Austrian acute myeloid leukemia study group. J Clin Oncol. 2011 Jul 1;29(19):2709-2716. doi: 10.1200/JCO.2011.35.0371

3. Ivey A, Hills RK, Simpson MA, et al: Assessment of minimal residual disease in standard-risk AML. N Engl J Med. 2016 Feb 4;374(5):422-433. doi: 10.1056/NEJMoa1507471

4. Shayegi N, Kramer M, Bornhauser M, et al: The level of residual disease based on mutant NPM1 is an independent prognostic factor for relapse and survival in AML. Blood. 2013 Jul 4;122(1):83-92. doi: 10.1182/blood-2012-10-461749

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

RNA is extracted from blood or bone marrow and reverse transcription is performed. Real time quantitative polymerase chain reaction (PCR) is performed from complementary DNA template using the LC480 instrument platform (Roche). This assay targets the most common recurrent NPM1 alterations in acute myeloid leukemia (A, B, and D insertion variants). The quantitative value of NPM1 messenger RNA copy number is determined relative to ABL1 as the reference transcript using the delta-delta CT method. The reproducible analytical sensitivity (limit of detection) of this part of the assay is approximately 0.01%.

 

DNA is extracted from blood or bone marrow and a PCR assay is performed using primers that amplify a fragment of NPM1 DNA containing the region susceptible to insertion variant. One of the PCR primers contains a fluorescent label. The amplified fragments are size separated by capillary electrophoresis. Wild type NPM1 produces a fragment length of 187-base pairs (bp). PCR fragments containing an insertional variant are observed as larger fragments, most typically 191-bp, as the majority of alterations are 4-bp insertions. The analytical sensitivity (limit of detection) of this part of the assay is approximately 5%.(Unpublished Mayo method)

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

Supplemental

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.

10 to 14 days

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

Blood/bone marrow: 2 weeks; Extracted DNA and RNA: 3 month

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.

81310-NPM1 (nucleophosmin) (eg, acute myeloid leukemia) gene analysis; exon 12 variants

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
NPM1Q NPM1 Mutation Analysis, V 54448-6
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.
MP053 Specimen Type 31208-2
605098 Interpretation 59466-3
605262 Signing Pathologist 19139-5

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