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Related Concept Videos

Real Time RT-PCR02:57

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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: May 29, 2025

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings
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Portable, quantitative, real-time isothermal nucleic acid amplification test using microfluidic device-coupled UV-LED

Natish Kumar1, Monika Kumari1, Devtulya Chander2

  • 1Department of Chemical Engineering, Indian Institute of Technology, Jammu, 181221, India.

Biosensors & Bioelectronics
|February 4, 2025
PubMed
Summary

A new Microfluidics Integrated LED-Photodiode (MILP) device enables rapid, portable nucleic acid testing at the point-of-care. This quantitative molecular diagnostic platform accurately detects SARS-CoV-2, offering a cost-effective solution for global health surveillance.

Keywords:
Isothermal nucleic acid testMicrofluidics-based portable detectorPhotovoltaic response analysisPoint of care diagnosticsQuantitative optical detectionReal-time analysis of disease load

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Area of Science:

  • Biomedical Engineering
  • Molecular Diagnostics
  • Point-of-Care Testing

Background:

  • Current molecular diagnostics often require complex laboratory infrastructure.
  • There is a need for rapid, portable, and quantitative nucleic acid testing solutions for point-of-care applications.
  • Existing point-of-care technologies may lack the sensitivity or specificity required for early disease detection.

Purpose of the Study:

  • To develop and validate a novel, automated, and portable Microfluidics Integrated LED-Photodiode (MILP) sensing technology.
  • To establish MILP as a quantitative molecular diagnostic platform for real-time nucleic acid testing.
  • To assess the performance of the MILP device for SARS-CoV-2 detection.

Main Methods:

  • Integration of a paper-based nucleic acid purification system with a polymer-based membrane filter.
  • In-situ isothermal amplification and dual-mode optical detection.
  • Quantitative signal analysis via photovoltaic response, electrical polarity, and photocurrent measurements.

Main Results:

  • The MILP device achieved an on-cartridge limit of detection of 10 copies/μL.
  • Demonstrated high clinical sensitivity (95%) and specificity (100%) for SARS-CoV-2 detection compared to real-time PCR.
  • Quantitative photovoltaic cut-offs identified test-gene-specific variations without auxiliary instrumentation.

Conclusions:

  • The MILP technology offers a stand-alone, fully quantitative, and portable solution for rapid molecular diagnostics.
  • This platform is suitable for intensive health surveillance, early disease screening, and monitoring in resource-limited settings.
  • The device eliminates the need for extensive and expensive laboratory infrastructure, facilitating widespread adoption.