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Related Experiment Video

Updated: Mar 9, 2026

Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
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Two-Stage Isothermal Enzymatic Amplification for Concurrent Multiplex Molecular Detection.

Jinzhao Song1, Changchun Liu1, Michael G Mauk1

  • 1Department of Mechanical Engineering and Applied Mechanics, School of Engineering and Applied Sciences, University of Pennsylvania, Philadelphia, PA.

Clinical Chemistry
|January 12, 2017
PubMed
Summary

A new rapid amplification (RAMP) assay enables highly multiplexed pathogen detection at the point-of-care. This innovative method provides sensitive and specific results in under 40 minutes, improving real-time diagnostics.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostics

Background:

  • Current single-plex tests struggle with the diverse range of infectious disease pathogens.
  • Multiplex PCR, while capable of detecting multiple targets, is confined to centralized labs, causing delays and limiting real-time diagnostic information.

Purpose of the Study:

  • To develop a point-of-care (POC) diagnostic assay for highly multiplexed pathogen detection.
  • To create a rapid, sensitive, and specific method for identifying multiple pathogens simultaneously.

Main Methods:

  • Proposed a novel 2-stage, nested-like, rapid isothermal amplification assay named rapid amplification (RAMP).
  • RAMP utilizes recombinase polymerase amplification (RPA) in the first stage for broad target amplification, followed by target-specific loop-mediated isothermal amplification (LAMP) in the second stage.
  • The assay is implemented on a microfluidic chip and detected using colorimetric or fluorescent dyes with smartphone compatibility.

Main Results:

  • RAMP demonstrated high multiplexing capability (≥16 targets) in both benchtop and microfluidic formats.
  • Achieved high sensitivity (detecting 1 plaque-forming unit of Zika virus) and specificity (no false positives or negatives).
  • The assay is rapid (<40 min), easy to use, and can process minimally prepared samples.

Conclusions:

  • RAMP is a hybrid, 2-stage assay combining RPA and LAMP advantages for rapid, sensitive, and specific multiplexed pathogen detection.
  • Its microfluidic format makes it suitable for point-of-care use, offering an inexpensive, on-site diagnostic tool for healthcare professionals.
  • RAMP addresses the limitations of existing methods by enabling real-time, comprehensive pathogen identification from a single sample.