NPM1 Quantitative for MRD

General Information

Lab Name

Lab Code

NPM1NG

Epic Ordering

NPM1 Insertion Mutation by NGS

Description

This quantitative test detects common insertion mutations in NPM1 gene and is intended for post-treatment minimal residual disease monitoring in NPM1-mutated AML. Purified genomic DNA is PCR amplified using double- indexed primers coupled with sequencing adaptors for subsequent next generation sequencing using an Illumina Miseq platform.

Reportable Range: 0.007 – 50.00%
Limit of Detection: 0.003%
Limit of Quantification: 0.007%
Precision near LOD (CV): 54%

Hematopathology test request form can be found at Request Form.

Forms & Requisitions

Requisition

Synonyms

Acute Myeloid Leukemia, AML, NPM, NPM-1, NPM1, NPM1 Exon 12 Insertion mutation, Nucleophosmin

Components

Code	Name
NPM1SD	NPM1 by NGS Specimen Descr.
NPM1RS	NPM1 by NGS Result
NPM1MP	NPM1 by NGS Mutation %
NPM1V	NPM1 by NGS Insertion Variant
NPM1IN	NPM1 by NGS Interpretation
NPM1M	NPM1 by NGS Method
NPM1GR	NPM1 by NGS Case Number

Interpretation

Method

PCR Amplification then Next Generation Sequencing

Reference Range

See individual components

Guidelines

Molecular Testing for Hematopoietic Disorders

Ordering & Collection

Specimen Type

Blood, Bone Marrow, Tissue - Fresh, frozen or FFPE/PET (Blocks), Fluids, Slides, Sorted Cells

Collection

Preferred:
Blood: 6mL in Lavender Top (EDTA)

Bone Marrow: 1-2mL in Lavender Top (EDTA)

Tissue, Fluids or Sorted cells: Sterile container in RPMI

Formalin-fixed paraffin embedded tissue (FFPE/PET) block

Slides: 10 unstained

Accepted:
Green Tops (Heparin), while green tops are accepted for testing, there is documentation that heparin can interfere with some PCR assays.

Extracted DNA from CLIA certified laboratory.

Formalin-fixed paraffin embedded tissue (FFPE/PET)

Tissue from curls cut from FFPE/PET blocks or stained cover-slipped slides

Unacceptable:
Specimens received beyond stability limit.
Specimens with low volume or inadequate WBC
Decalcified or unacceptable fixatives

Forms & Requisitions

Requisition

Handling Instructions

Deliver samples to Hematopathology as soon as possible.
If sample is older than 48 hours, please store at 2-6°C upon receipt and send refrigerated.
Samples stored at room temperature or refrigerated older than 7 days will be rejected.
Frozen samples should be shipped on dry ice.

Quantity

requested: Blood: 6mL EDTA/BMA: 1-2mL EDTA/Tissue Blocks: Adequate size to cut 10-20 curls/Slides: 10 unstained
minimum: Blood: 1mL EDTA/ BMA: 0.5mL EDTA/Tissue Blocks: 5 curls/Slides: 5 slides

Processing

Processing: Send specimens to Hematopathology – Molecular department for processing.
The stability limit for Blood and Bone Marrow is 7 days, samples beyond this limit will be rejected.

Specimens must be received in lab by 7am on Wednesday to be included on run.

Performance

LIS Dept Code

Molecular Hematopathology (MHP)

Performing Location(s)

Other	Hematopathology, Molecular 206-606-7060 UW Hematopathology Laboratory, G7-800 Fred Hutchinson Cancer Center 825 Eastlake Avenue E. Seattle, WA 98109	Hematopathology, Molecular hours: 7:00 am - 5:30 pm M-F 8:00 am - 4:30 pm Saturday, closed on Sunday, and with reduced staffing on major holidays

Frequency

Performed once weekly on Wednesdays; results may take up to 10 days.

Available STAT?

Billing & Coding

CPT codes: 81310
LOINC: 54448-6
Interfaced Order Code: UOW3029