Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

67.6K
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...
67.6K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Challenges to initiate and complete a 4-month rifapentine-moxifloxacin TB treatment regimen.

The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease·2025
Same author

Final Search for Short-Baseline Neutrino Oscillations with the PROSPECT-I Detector at HFIR.

Physical review letters·2025
Same author

Retraction Note: Glypican-3 reexpression regulates apoptosis in murine adenocarcinoma mammary cells modulating PI3K/Akt and p38MAPK signaling pathways.

Breast cancer research and treatment·2023
Same author

Final Measurement of the ^{235}U Antineutrino Energy Spectrum with the PROSPECT-I Detector at HFIR.

Physical review letters·2023
Same author

Mortality-Air Pollution Associations in Low Exposure Environments (MAPLE): Phase 2.

Research report (Health Effects Institute)·2022
Same author

Genomic loci associated with performance limiting equine overriding spinous processes (kissing spines).

Research in veterinary science·2022

Related Experiment Video

Updated: Apr 17, 2026

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications
07:40

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications

Published on: June 29, 2020

14.8K

Insulated Isothermal Reverse Transcriptase PCR (iiRT-PCR) for Rapid and Sensitive Detection of Classical Swine Fever

O Lung1, J Pasick2, M Fisher2

  • 1National Centres for Animal Disease, Lethbridge Laboratory, Canadian Food Inspection Agency, Lethbridge, AB, Canada.

Transboundary and Emerging Diseases
|February 4, 2015
PubMed
Summary

A new insulated isothermal reverse transcriptase PCR (iiRT-PCR) assay rapidly detects classical swine fever virus (CSFV) using a user-friendly device. This sensitive diagnostic tool aids in early disease surveillance and outbreak monitoring for classical swine fever.

Keywords:
PCRPoint-of-careclassical swine fever virusisothermal amplificationpestivirus

More Related Videos

Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
08:10

Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses

Published on: May 2, 2019

9.0K
Rapid Diagnosis of Avian Influenza Virus in Wild Birds: Use of a Portable rRT-PCR and Freeze-dried Reagents in the Field
08:46

Rapid Diagnosis of Avian Influenza Virus in Wild Birds: Use of a Portable rRT-PCR and Freeze-dried Reagents in the Field

Published on: August 2, 2011

16.2K

Related Experiment Videos

Last Updated: Apr 17, 2026

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications
07:40

Field Postmortem Rabies Rapid Immunochromatographic Diagnostic Test for Resource-Limited Settings with Further Molecular Applications

Published on: June 29, 2020

14.8K
Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
08:10

Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses

Published on: May 2, 2019

9.0K
Rapid Diagnosis of Avian Influenza Virus in Wild Birds: Use of a Portable rRT-PCR and Freeze-dried Reagents in the Field
08:46

Rapid Diagnosis of Avian Influenza Virus in Wild Birds: Use of a Portable rRT-PCR and Freeze-dried Reagents in the Field

Published on: August 2, 2011

16.2K

Area of Science:

  • Veterinary Virology
  • Molecular Diagnostics
  • OIE-listed Diseases

Background:

  • Classical swine fever (CSF) poses a significant threat to the swine industry globally.
  • Current diagnostic methods may lack the speed, sensitivity, or field-deployability required for effective surveillance and outbreak response.
  • Development of user-friendly, rapid, and sensitive diagnostic tools is crucial for managing CSF.

Purpose of the Study:

  • To validate a novel probe-based insulated isothermal reverse transcriptase PCR (iiRT-PCR) assay for the rapid detection of classical swine fever virus (CSFV).
  • To evaluate the performance of the assay on a compact, user-friendly nucleic acid analyzer for field applications.
  • To assess the assay's sensitivity, specificity, and turnaround time for CSFV diagnosis.

Main Methods:

  • Validation of a probe-based iiRT-PCR assay targeting CSFV.
  • Testing the assay on a diverse panel of 33 CSFV strains and 18 other livestock-associated viruses for specificity.
  • Assessing detection limits (LOD95%) and early detection in experimentally infected pigs.
  • Evaluating the turnaround time from sample to result using extracted RNA and serum samples.

Main Results:

  • The iiRT-PCR assay accurately detected all 33 CSFV strains tested, including all three genotypes and a vaccine strain.
  • No cross-reactivity was observed with a comprehensive panel of 18 other relevant livestock viruses, including other pestiviruses and African swine fever virus.
  • The assay detected CSFV RNA as early as 2 days post-inoculation in experimentally infected pigs, prior to clinical signs.
  • The limit of detection (LOD95%) was determined to be 23 copies per reaction.
  • Sample-to-answer turnaround time was under one hour using extracted RNA or serum.

Conclusions:

  • The validated iiRT-PCR assay is a sensitive and specific tool for rapid CSFV detection.
  • The user-friendly, compact device facilitates automated data interpretation, making it suitable for field deployment.
  • This diagnostic approach offers a valuable tool for enhanced surveillance and outbreak monitoring of classical swine fever.