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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Related Experiment Video

Updated: Mar 15, 2026

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings
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Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification.

Michael G Mauk1, Changchun Liu2, Jinzhao Song3

  • 1School of Engineering and Applied Sciences, University of Pennsylvania, 220 S 33 rd Street, Philadelphia, PA 19104, USA. mmauk@seas.upenn.edu.

Microarrays (Basel, Switzerland)
|September 8, 2016
PubMed
Summary

This study presents a portable microfluidic chip for rapid, low-cost nucleic acid amplification tests (NAATs) suitable for point-of-care diagnostics. The "lab on a chip" device enables sensitive HIV viral load quantification comparable to lab-based PCR.

Keywords:
isothermal nucleic acid amplificationlab on a chipmicrofluidics

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Area of Science:

  • Biomedical Engineering
  • Molecular Diagnostics
  • Microfluidics

Background:

  • Conventional nucleic acid amplification tests (NAATs) often require centralized laboratory facilities, limiting their use in point-of-care (POC) settings.
  • Developing rapid, field-deployable NAATs is crucial for timely disease diagnosis and monitoring.

Purpose of the Study:

  • To describe microfluidic components and systems for rapid, low-cost, field-deployable NAATs.
  • To demonstrate a portable POC NAAT for HIV viral load quantification using a "lab on a chip" approach.

Main Methods:

  • Nucleic acids (NAs) are extracted from blood samples using an embedded membrane within a disposable microfluidic chip.
  • Isothermal amplification of captured NAs is performed at ~65 °C, with real-time fluorescence monitoring via a smartphone camera.
  • Pre-stored, temperature-activated lyophilized reagents are utilized for simplified assay execution.

Main Results:

  • Achieved Limits of Detection (LOD) better than 10³ virons/sample.
  • Demonstrated a portable POC NAAT with performance comparable to conventional polymerase-chain reaction (PCR) tests.
  • Developed a companion device for centrifuge-free plasma extraction from whole blood.

Conclusions:

  • A portable, miniaturized "lab on a chip" NAAT device is feasible for rapid, field-deployable molecular diagnostics.
  • The technology offers a low-cost, convenient alternative to traditional laboratory-based NAATs.
  • Potential applications extend to food testing, cancer screening, and insect genotyping.