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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

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: Jun 28, 2026

Using a Pan-Viral Microarray Assay (Virochip) to Screen Clinical Samples for Viral Pathogens
13:45

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Real-time microchip PCR for detecting single-base differences in viral and human DNA

M S Ibrahim1, R S Lofts, P B Jahrling

  • 1United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland 21702, USA. sibrahim@detrick.army.mil

Analytical Chemistry
|May 26, 1998
PubMed
Summary
This summary is machine-generated.

This study introduces a portable device for rapid genetic analysis, enabling quick identification of single-base differences in viral and human DNA. This technology accelerates infectious disease and genetic disorder diagnosis using real-time PCR assays.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Virology

Background:

  • Rapid and accurate identification of single-nucleotide polymorphisms (SNPs) is crucial for diagnosing infectious diseases and genetic disorders.
  • Existing diagnostic methods can be time-consuming and require specialized laboratory equipment.

Purpose of the Study:

  • To develop and validate real-time 5' nuclease PCR assays for distinguishing single-base polymorphisms.
  • To demonstrate the feasibility of using a battery-powered miniature analytical thermal cycling instrument (MATCI) for rapid genetic analysis.

Main Methods:

  • Development of consensus Orthopoxvirus PCR primers targeting the hemagglutinin (HA) gene.
  • Design of vaccinia virus-specific fluorogenic (TaqMan) probes for single-nucleotide substitution detection.
  • PCR amplification of a human complement component C6 gene segment and TaqMan probe-based polymorphism detection.
  • Utilized a rapid DNA preparation method for streamlined sample processing.

Main Results:

  • The MATCI successfully distinguished single-base differences in both Orthopoxvirus DNA and human genomic DNA.
  • The developed PCR assays accurately identified specific nucleotide substitutions within the HA gene and C6 gene.
  • Achieved complete workflow from sample to real-time detection in under one hour.

Conclusions:

  • The MATCI is a feasible instrument for rapid diagnosis of infectious diseases and genetic disorders.
  • Real-time 5' nuclease PCR assays coupled with the MATCI offer a fast and accurate method for single-base polymorphism detection.
  • This technology has the potential to significantly improve point-of-care diagnostics.