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The Invader assay for SNP genotyping.

Michael Olivier1

  • 1Human and Molecular Genetics Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA. moliver@mcw.edu

Mutation Research
|April 15, 2005
PubMed
Summary

The Invader assay utilizes flap endonuclease (FEN) for precise single nucleotide polymorphism (SNP) genotyping. This method offers high specificity and sensitivity for DNA analysis, with potential for high-throughput applications.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The Invader assay is a biochemical method for detecting single nucleotide polymorphisms (SNPs).
  • It relies on the enzymatic activity of flap endonucleases (FENs) to cleave specific DNA structures.
  • SNP genotyping is crucial for various applications, including disease association studies and personalized medicine.

Purpose of the Study:

  • To describe the mechanism and detection methods of the Invader assay.
  • To highlight the specificity, sensitivity, and potential applications of the Invader assay.
  • To discuss advancements enabling multiplexing and high-throughput SNP genotyping.

Main Methods:

  • The assay employs allele-specific overlapping oligonucleotides that hybridize to target DNA containing a SNP.
  • A structure-specific flap endonuclease (FEN), termed cleavase, cleaves the hybridized oligonucleotide upon correct allele matching.
  • Detection of cleavage can be achieved through fluorescence resonance energy transfer (FRET) cassettes, fluorescence polarization (FP) probes, or mass spectrometry.

Main Results:

  • The Invader assay demonstrates high specificity and a low failure rate in SNP detection.
  • The assay is capable of detecting zeptamol quantities of target DNA, indicating high sensitivity.
  • Novel chip- or bead-based approaches are being developed for multiplexing and high-throughput SNP genotyping.

Conclusions:

  • The Invader assay is a robust and sensitive method for SNP genotyping.
  • Its enzymatic cleavage mechanism ensures high accuracy.
  • Adaptations for multiplexing and high-throughput analysis position the Invader assay for broader applications in genetic research and diagnostics.

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