Benefits & Overview

MSI Markers Covered 

Workflow

Specifications

Request More Information

Key Features

Sensitive

Limit of detection down to 1% VAF

User Friendly

Matched normal not required

Included software reports MSI-H, MSI-L/MSS

Fast Turnaround

Results in less than two hours

Batch up to 46 samples per qPCR run

Versatile

Compatible with tumor tissue or cell-free DNA (cfDNA)

Overview

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

The VarTrace Microsatellite Instability (MSI) Assay enables highly sensitive detection of MSI. This assay is intended for the qualitative detection of mutations in five MSI loci without the need of matched normal tissue.

NuProbe’s VarTrace MSI Assay utilizes patented BDA technology to selectively amplify the alleles from MMR deficient samples enabling detection of MSI-high (MSI-H) status with a limit of detection to 1% variant allele frequency (VAF). The assay interrogates five mononucleotide markers (BAT-25, BAT-26, NR-21, NR-24, and MONO-27) to determine MSI status for each sample.

Request More info

Microsatellite Instability Markers Covered

BAT-25, BAT-26, NR-21, NR-24, and MONO-27

Sample Data

BAT26 MSI-H
  • Limit of Detection Performance
  • VarTrace MSI assay shows superb analytical limit of detection down to 1% allele frequency of the MSI unstable molecules.
  • MSI Determination
  • The MSI status is based on the Ct values for each marker in the sample, by comparing with the GAPDH internal control to normalize the sample input. If the Ct value of marker is earlier than threshold, the marker is a positive. And based on the number of positive markers out of the 5 markers, we can call the sample MSI-H (>2/5); MSI-L(1/5) or MSS(0/5). Figure is based on hypothetical data.
  • MSI Software
  • We provide an easy-to-use software to interpret MSI results and generate a report. Simply upload the qPCR data into the NuProbe MSI software, and the software gives sample results in a plate view map by color code. Users can then label the samples, view the result for each marker and qPCR curves, and export the MSI status report in Excel format.
MSI-H vs MSS
  • qPCR Data Curve
  • qPCR curve results from a colorectal cancer MSS sample and a colorectal cancer MSI sample.

Workflow

Step 1: DNA Extraction

DNA is extracted from clinical samples with commercial extraction kits such as cobas® cfDNA Sample Preparation Kit, QIAamp® DNA Blood Mini Kit, GeneRead® DNA FFPE Kit.

Sanger Sequencing Workflow

Step 2: qPCR BDA Reaction

NuProbe’s qPCR kit reagents and clinical samples are prepared for qPCR reactions. After running the reaction, Ct results are obtained.

1.5 hrs (15 min hands-on)

Computer Analysis Icon

Step 3: Results Interpretation

The MSI status of each sample will be visualized and reported by the easy-to-use VarTrace MSI software.

2 min hands-on

Download Product User Manual

    Compatible Instruments

    BioRad CFX96

    BioRad CFX96

    The assay has been optimized with BioRad CFX96. To use other qPCR instruments, please contact us.

    Specifications

    NuProbe Kit Box
    Catalog Number

    1R-0401-S
    48 Reactions

    Technologymulti-blocker mBDA & qPCR
    Kit size48 specimens / kit
    BiomarkersTube 1: BAT26, NR21, NR24
    Tube 2: BAT25, Mono27
    Internal ControlGAPDH
    Tubes/wells per specimen2
    Turnaround timeqPCR only: 1h30min including approximately 15min hands-on time
    Limit of detection≥ 10ng per reaction: 1%
    < 10ng per reaction: 5%
    Sensitivity>98%
    Specificity>98% at 1% VAF
    Specimen typeBlood or FFPE
    Specimen input1.5-15 ng per reaction (3-30 ng per specimen) ideally 20ng (10 ng per tube)
    ControlPositive (5%) and Negative
    Amplicon length[80, 120] bp
    Storage condition-20°C, up to 6 months
    InstrumentsBioRad CFX96
    SoftwareIncluded software reports MSS, MSI-L, MSI-H based on N/5 abnormal

    Request More Info

    Alessandro Pinto, Ph.D., Director of Service and Partnership at NuProbe.

    Fill out the form to contact Ale for questions about the VarTrace products.

    Ale