Key Features

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Sensitive

Limit of detection down to 0.1% VAF

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Quantitative

Able to quantify mutation VAF in original sample

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Comprehensive

Wide range of mutations covered

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User-friendly

Easy workflow

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Benefits

  • Detects 254 AML hotspot mutations in 6 genes: FLT3, DNMT3A, IDH1, IDH2, KIT, and NPM1
  • Requires 80 ng – 1.5 μg of DNA from whole blood sample
  • Delivers results in 8 hours, with 2.5 hours of hands-on time

Sample Data

Product AML Sample Data

Variant verification by Sanger sequencing

Because NuProbe’s BDA technology enriches the variant allele frequency (VAF) of mutations by over 200-fold, Sanger’s limit of detection of 20% VAF is effectively improved to 0.1% VAF. Sanger sequencing can be applied to confirm the identity of quantities detected and quantitated by qPCR using standard sequencing protocols.

Workflow

Workflow Step 1

Step 1: DNA Extraction

DNA is extracted from clinical samples with commercial extraction kits such as QIAamp® DNA Blood Mini Kit.
40 min (40 min hands-on)

Sanger Sequencing Workflow

Step 2: qPCR BDA Reaction

NuProbe’s qPCR/Sanger kit reagents and clinical samples are prepared for qPCR reactions. After running the reaction, Ct results and BDA products are obtained.
1.5 hrs (30 min hands-on)

Workflow Step 3

Step 3: Sanger Sequencing

BDA products are prepared for Sanger sequencing, including PCR product purification, cycle sequencing and sequencing clean-up.
3.5 hrs (1hr hands-on)

Purified Sanger products are analyzed by capillary electrophoresis.
2 hrs (15 min hands-on)

Compatible Instruments

qPCR

qPCR

BioRad CFX96


qPCR

ABI 7500


Cobas z480

Cobas z480

Sanger sequencing

Sanger Sequencing

ABI 3500 Dx Genetic Analyzer

Background

For research use only. Not for use in diagnostic procedures.

In patients with Acute Myeloid Leukemia (AML), several genes are frequently mutated, including FLT3, IDH1, IDH2, DNMT3A, KIT, and NPM1. The presence of these specific mutations can guide treatment with targeted therapies such as RYDAPT® (midostaurin), XOSPATA® (gilteritinib), TIBSOVO® (ivosidenib), and IDHIFA® (enasidenib).

The VarTrace™ AML Panel enables highly sensitive detection and quantification of mutations for AML patients. The kit uses NuProbe’s PCR-based Blocker Displacement Amplification (BDA) technology to enable the selective amplification of low abundant sequence variants (SNV and indels) in a background of wildtype DNA. Following PCR enrichment, Sanger sequencing is applied to reveal the identity of the variants.

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