FLT3 & NPM1 Mutation Detection, Whole Blood or Bone Marrow Aspirate

FLT3 & NPM1 Mutation Detection

Order Code:	FLT3/NPM1
Order Form:	Molecular Oncology Requisition

Specimen:

Whole Blood or Bone Marrow Aspirate

Collection Medium:

Lavender top tube 3 mL (EDTA)

Minimum:

3 mL of peripheral blood or bone marrow aspirate. Specimens for which the AML blast count is at least 10% will be tested. Testing on smaller volumes may be attempted. However, this may compromise the ability to perform testing. Testing requires a dedicated collection tube.

Testing Schedule:

0800-1700 Testing offered once per week excluding weekends and University holidays.

Turn Around Time:

7 working days

Reference Range:

Expected PCR fragment sizes for each DNA target are provided below for unaffected and affected samples. The vast majority of affected persons harboring one or more of these mutations are heterozygous, meaning both products are present at the diagnostic locus. Unaffected Affected (negative) (positive) FLT3-ITD: 330 bp >330 bp FLT3-D835: 67 bp 197 bp NPM1: 426 bp 430 bp

Comments:

FLT3 and NPM1 Mutation Detection Assays test for two different mutations in the FLT3 gene and an insertion in exon 12 of the NPM1 gene, all of which are somatic mutations associated with acute myelogenous leukemia (AML). These molecular markers are useful in diagnosis and clinical stratification of AML patients. Detection of FLT3 in-frame internal tandem duplications (FLT3-ITD mutation) is performed as a mail-out test (sent to a reference laboratory), whereas the assays that detect FLT3 point mutations that alter the aspartic acid at position 835 (FLT3-D835) and the NPM1 mutations are performed in the University of Iowa Molecular Pathology Laboratory. To avoid confusion, the results for all 3 targets are reported together in a consolidated interpretation.

Methodology:

Multiplex PCR followed by Eco RV digestion with fluorescent fragment analysis on the ABI 3130 (capillary electrophoresis).

Instructions:

Testing is generally not available on a STAT basis. However, expedited testing can be arranged by contacting the Molecular Pathology Laboratory at 384-9568 or the Molecular Genetic Pathology Fellow (pager #8682).

Sample Processing:

Label transport tube with two patient identifiers, date and time of collection.
Patient's age and sex is required on requisition for processing.
Relevant clinical information must be submitted with specimen in order to provide correct interpretation of test results.
Lavender tube is sent as whole blood, no processing necessary.
Do Not Centrifuge.

Sample Storage:

Room temperature for up to 24 hours, then refrigerate the whole blood if it is necessary to be held overnight, or weekends, or holidays.

Transport Instructions:

Place requisition into outside pocket of bag.
Recommend express mail or equivalent if not on courier service.

CPT Code:

83890, 83892 x2, 83898 x3, 83900, 83909 x2, 83912