Specimen

Whole Blood or Bone Marrow Aspirate

Collection Medium:

Minimum:

Adults 3 ml whole blood.  Testing on smaller volumes than those
requested will be attempted. However, in some cases, small blood
volumes may compromise the ability to perform testing.  Testing
requires a dedicated collection tube.

For instructions on submission of bone marrow specimens, please call 
the Molecular Pathology Laboratory at 319-384-9568.

Delivery 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.

Testing Schedule:

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

Analytic 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:     329 bp          >329 bp
 FLT3-D835:     67 bp           197 bp
 NPM1:         426 bp           430 bp

Comments:

This is a qualitative assay that simultaneously detects the following 
mutations: FLT3-ITD (internal tandem duplication in exons 14-15 of the 
FLT3 gene), FLT3 D835 (point mutations resulting in a substitution at 
position Asp835 for FLT3 tyrosine kinase and loss of Eco RV restriction 
site), and internal 4 bp duplication/insertion in exon 12 of 
nucleophosmin (NPM1) gene, all of which serve as molecular markers 
useful in diagnosis and clinical stratification of acute myeloid 
leukemia (AML).

Methodology:

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

CPT Code:

83912, 83890, 83900, 83901, 83892, 83909