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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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Author Spotlight: Advancements in Multiplex Detection of Respiratory Viruses
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Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination.

Kuan-Lun Ho1, Jing Ding1, Jia-Shao Fan2

  • 1Department of Mechanical and Nuclear Engineering, Kansas State University, Manhattan, KS 66506, USA.

Micromachines
|August 26, 2023
PubMed
Summary
This summary is machine-generated.

A new digital microfluidics (DMF) platform using reverse transcription quantitative polymerase chain reaction (RT-qPCR) can detect and differentiate SARS-CoV-2 wildtype, Delta, and Omicron variants rapidly. This multitarget detection aids in tracking and reducing COVID-19 transmission.

Keywords:
COVID-19Delta variantOmicron variantRT-qPCRSARS-CoV-2digital microfluidicselectrowetting

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Area of Science:

  • Virology
  • Molecular Biology
  • Biotechnology

Background:

  • Continuous SARS-CoV-2 mutations have increased transmissibility, prolonging the COVID-19 pandemic and straining healthcare systems.
  • The emergence of variants of concern (VOCs) like Delta and Omicron necessitates rapid detection and differentiation for effective disease control.
  • A point-of-care (POC) platform for multitarget detection is crucial for real-time monitoring and management of viral spread.

Purpose of the Study:

  • To develop and evaluate a digital microfluidics (DMF)-based cartridge for rapid, multitarget detection of SARS-CoV-2 genotypes.
  • To discriminate between the wildtype, Delta, and Omicron variants of SARS-CoV-2 using reverse transcription quantitative polymerase chain reaction (RT-qPCR).
  • To assess the diagnostic capability of the DMF RT-qPCR platform for clinical applications.

Main Methods:

  • Utilized a digital microfluidics (DMF) platform integrated with reverse transcription quantitative polymerase chain reaction (RT-qPCR).
  • Developed a multitarget assay capable of detecting and differentiating SARS-CoV-2 wildtype, Delta, and Omicron variants.
  • Evaluated performance using clinical analog samples spiked with various SARS-CoV-2 strains and 18S rDNA for internal control.

Main Results:

  • The DMF RT-qPCR platform demonstrated high sensitivity (10 copies/μL) and amplification efficiency (96.1%) when tested with Omicron variant RNA samples.
  • Accurate detection and discrimination of wildtype, Delta, and Omicron SARS-CoV-2 strains were achieved in clinical analog samples within 49 minutes.
  • The platform successfully identified different SARS-CoV-2 strains, indicating its potential for rapid clinical diagnosis.

Conclusions:

  • The developed DMF RT-qPCR platform offers a sensitive and specific solution for detecting and differentiating SARS-CoV-2 variants at the point-of-care.
  • This technology can significantly aid in tracking viral spread, managing outbreaks, and informing public health strategies.
  • The rapid multitarget detection capability of this platform is vital for controlling the ongoing COVID-19 pandemic.