Contributions
Abstract: PB1689
Type: Publication Only
Background
Acute myeloid leukemia (AML) in general has a poor prognosis. Assessment of the mutation status of the FLT3 (fms related tyrosine kinase 3) receptor gene in AML is the most important prognostic indicator of disease outcome, which is often substantial, as many studies in AML have shown that the presence of FLT3 activating mutations portends a poor prognosis. The LeukoStrat® CDx FLT3 Mutation Assay targets regions of the FLT3 gene to identify internal tandem duplication (ITD) mutations and tyrosine kinase domain (TKD) mutations. Since this assay is a signal ratio (SR) assay with a validated cutoff of 0.05, demonstration of international harmonization of results is paramount.
Aims
To assess the performance of the Invivoscribe® LeukoStrat® CDx FLT3 Mutation Assay.
Methods
Results
The analytical performance of the LeukoStrat® CDx FLT3 Mutation Assay was evaluated using contrived LDB samples, with known FLT3 mutations. For limit of blank (LoB), the SR was 0.00 in the ITD assay and 0.00 to 0.01 in the TKD assay, which is well below the clinical cutoff SR of 0.05. The limit of detection in the ITD assay detected allelic ratios of 0.03, 0.05, and 0.33 above the LoB SR in more than 95% of samples for insertions sized at 30 bp, 126 bp and 279 bp, respectively. The limit of detection in the TKD assay detected an allelic ratio of 0.05 above the LoB. For precision and reproducibility, the SR %CV was within 3-14% across ITD and TKD mutation types regardless of reagent lots, equipment, or operator.
Conclusion
This robust assay produced a SR %CV less than 15% regardless of reagent lot, equipment or operator. The high reproducibility between the three laboratories on three different continents provides evidence that the Invivoscribe® LeukoStrat® CDx FLT3 Mutation Assay is an internationally standardized assay.
Session topic: 4. Acute myeloid leukemia - Clinical
Keyword(s): Tyrosine kinase, Flt3-ITD, Acute Myeloid Leukemia
Abstract: PB1689
Type: Publication Only
Background
Acute myeloid leukemia (AML) in general has a poor prognosis. Assessment of the mutation status of the FLT3 (fms related tyrosine kinase 3) receptor gene in AML is the most important prognostic indicator of disease outcome, which is often substantial, as many studies in AML have shown that the presence of FLT3 activating mutations portends a poor prognosis. The LeukoStrat® CDx FLT3 Mutation Assay targets regions of the FLT3 gene to identify internal tandem duplication (ITD) mutations and tyrosine kinase domain (TKD) mutations. Since this assay is a signal ratio (SR) assay with a validated cutoff of 0.05, demonstration of international harmonization of results is paramount.
Aims
To assess the performance of the Invivoscribe® LeukoStrat® CDx FLT3 Mutation Assay.
Methods
Results
The analytical performance of the LeukoStrat® CDx FLT3 Mutation Assay was evaluated using contrived LDB samples, with known FLT3 mutations. For limit of blank (LoB), the SR was 0.00 in the ITD assay and 0.00 to 0.01 in the TKD assay, which is well below the clinical cutoff SR of 0.05. The limit of detection in the ITD assay detected allelic ratios of 0.03, 0.05, and 0.33 above the LoB SR in more than 95% of samples for insertions sized at 30 bp, 126 bp and 279 bp, respectively. The limit of detection in the TKD assay detected an allelic ratio of 0.05 above the LoB. For precision and reproducibility, the SR %CV was within 3-14% across ITD and TKD mutation types regardless of reagent lots, equipment, or operator.
Conclusion
This robust assay produced a SR %CV less than 15% regardless of reagent lot, equipment or operator. The high reproducibility between the three laboratories on three different continents provides evidence that the Invivoscribe® LeukoStrat® CDx FLT3 Mutation Assay is an internationally standardized assay.
Session topic: 4. Acute myeloid leukemia - Clinical
Keyword(s): Tyrosine kinase, Flt3-ITD, Acute Myeloid Leukemia