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Related Experiment Video

Updated: May 17, 2026

Visual Detection of Multiple Nucleic Acids in a Capillary Array
08:56

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Published on: November 15, 2017

IsoPCR: an analytically sensitive, nested, multiplex nucleic acid amplification method.

Martin Jensen Søe1, Mikkel Rohde, Jens Mikkelsen

  • 1Atonomics A/S, Copenhagen, Denmark. mjs@atonomics.com

Clinical Chemistry
|November 2, 2012
PubMed
Summary
This summary is machine-generated.

The novel isoPCR method offers rapid, highly specific multiplex nucleic acid detection. This molecular diagnostic tool achieves a limit of detection down to 1 copy, significantly improving upon existing methods.

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Area of Science:

  • Molecular Biology
  • Nucleic Acid Amplification
  • Diagnostic Development

Background:

  • Simultaneous multi-target nucleic acid detection requires high sensitivity, specificity, and speed.
  • Existing methods face limitations in achieving these combined attributes.
  • The iso-structure PCR (isoPCR) method was developed to address these needs.

Purpose of the Study:

  • To introduce and evaluate the novel isoPCR method for nucleic acid detection.
  • To compare isoPCR's performance against established methods like nested quantitative PCR (qPCR) and loop-mediated isothermal amplification (LAMP).
  • To assess isoPCR's multiplexing capability for detecting low-copy number pathogens.

Main Methods:

  • IsoPCR employs a two-stage, nested-like amplification process: multiplex preamplification followed by isothermal amplification.
  • Performance was evaluated for detecting Candida glabrata DNA.
  • Multiplexing capability was tested for simultaneous detection of four sepsis-associated pathogens.

Main Results:

  • IsoPCR achieved a limit of detection (LOD) of 1 copy for Candida glabrata, 5-fold lower than nested qPCR (5 copies) and significantly better than LAMP assays.
  • Amplification time was halved compared to nested qPCR.
  • IsoPCR demonstrated high theoretical specificity by recognizing 6 regions versus 4 for nested qPCR.
  • Simultaneous detection of 4 pathogens with LODs ≤10 copies was successful, showcasing multiplexing and specificity in complex samples.

Conclusions:

  • IsoPCR is a powerful molecular diagnostic tool for multiplex nucleic acid detection.
  • It offers a low limit of detection (down to 1 copy) and high theoretical specificity.
  • The method significantly reduces amplification time compared to nested qPCR.