HemaMut Human FLT3-ITD Mutation Quantitative Detection Kit (Fragment Analysis)

Background

FLT3 mutation is predominantly found in primary AML and a few in ALL. Internal tandem duplication (ITD) and tyrosine kinase domain (TKD) point mutations are two common mutation types. FLT3-ITD mutations occur in approximately 30% of AML patients, more common than FLT3-TKD mutations, which occur in approximately 10% of patients.

The constitutive activation of FLT3 by mutated FLT3-ITD is one of the most common molecular changes in AML. According to the NCCN-AML guidelines, wild-type NPM1 with FLT3-ITDhigh indicates an adverse prognosis; wild-type NPM1 with FLT3-ITDlow indicates an intermediate prognosis in AML patients. For AML patients with FLT3-ITD mutation, kinase inhibitors such as sorafenib, midostaurin, and sunitinib can be used for combination therapy.

Features

Simple operation and less time consumption; reliable results and intuitive interpretation; high sensitivity and sample saving.

Detection Process

DNA is extracted from peripheral blood or bone marrow blood;

1) Fluorescent PCR primers covering the JM domain are mixed with target DNA for PCR amplification reactions.
2) The PCR amplified products are analyzed by capillary electrophoresis using an ABI instrument. Result analysis: ITD mutant is determined by PCR product size. The wild-type product is 383 bp. FLT3-ITD mutation has a longer PCR product than the wild-type product ( > 383 bp). Mutation type (Figure 1).

Figure 1

Kit Detection Results

Amplified products are subjected to capillary electrophoresis on an ABI genetic analyzer and analyzed with GeneMarker software.

The analysis results of positive samples are shown in the figure below:

Figure 2. Positive results

The analysis results of negative samples are shown in the figure below:

Figure 3. Negative results

Specimen Requirements

Peripheral blood or bone marrow (3–5 mL) is collected in the EDTA tube and stored at 2–8 °C for not more than 5 days. RNA extraction is performed within 5 days.

References

[1] Blood. 2002;100:4372-4380.

[2] Blood.2008;111:1552-1559.

[3] Lancet. 2006;368(9550):1894-1907.

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