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Test Code LAB9765 Janus Kinase 2 (JAK2 V617F) Mutation Assay

Methodology

The assay detects the JAK2 V617F/G1849T mutation using PCR primers and allele-specific dye-labeled oligonucleotides that exactly match the mutant or normal alleles. When perfectly hybridized to the template DNA sequence, the labeled oligonucleotides are cleaved by the 5’ -> 3’ exonuclease activity of DNA polymerase. Cleavage during the exponential phase of PCR releases the reporter dye from the quencher and the signal is quantified ("real time quantitative PCR"). The results report the mutant allele proportion of total JAK2 alleles.


The validated limit of detection for this assay is 0.1%.

Performing Laboratory

BJH Molecular Diagnostic Laboratory

Specimen Requirements

Call Barnes-Jewish Hospital Molecular Diagnostics Laboratory at 314-454-8685 for collection instructions.

 

Form: Molecular Diagnostics Requisition

 

Submit only 1 of the following specimens:

 

Specimen Type: Blood

Container/Tube: Lavender top (EDTA)

Specimen Volume: Full tube

Specimen Minimum Volume: 1 to 2 mL

Collection Instructions:

1. Invert several times to mix blood. Clotted blood is not acceptable.

2. Forward promptly at ambient temperature only. Specimen cannot be frozen.

 

Specimen Type: Bone marrow

Container/Tube: Lavender top (EDTA)

Specimen Volume: Full tube

Specimen Minimum Volume: 1 to 2 mL

Collection Instructions:

1. Invert several times to mix bone marrow. Clotted bone marrow is not acceptable.

2. Forward promptly at ambient temperature only. Specimen cannot be frozen.

Specimen Transport Temperature

Ambient/Refrigerate/Frozen NO

Reference Values

An interpretative report will be provided.

Day(s) Test Set Up

Tuesday
Turnaround Time:
STAT: not available
Routine: 5 to 7 business days

Test Classification and CPT Coding

81270 

Additional Information

For BJH Laboratory Use Only

Analyte Stability:
Blood and bone marrow is stable 0 to 4 hours at ambient temperature, up to 168 hours at 4° C.