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 Experiment Videos

A new method for measuring reverse transcriptase activity by ELISA.

J Eberle1, R Seibl

  • 1Max von Pettenkofer-Institut, Ludwig-Maximilians-University of München, Germany.

Journal of Virological Methods
|December 1, 1992
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Severe underquantification of HIV-1 group O isolates by major commercial PCR-based assays.

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases·2020
Same author

Transmission of HIV Drug Resistance and the Predicted Effect on Current First-line Regimens in Europe.

Clinical infectious diseases : an official publication of the Infectious Diseases Society of America·2015
Same author

Asymmetry in genitalia does not increase the rate of their evolution.

Molecular phylogenetics and evolution·2015
Same author

Age-related changes in auditory nerve-inner hair cell connections, hair cell numbers, auditory brain stem response and gap detection in UM-HET4 mice.

Neuroscience·2015
Same author

Haemophilia-related outcome after liver transplantation and treatment with sofosbuvir/ribavirin in a HCV-HIV coinfected man with liver failure and hepatocellular carcinoma.

Haemophilia : the official journal of the World Federation of Hemophilia·2015
Same author

Interleukin-33 promotes the proliferation of mouse mast cells through ST2/MyD88 and p38 MAPK-dependent and Kit-independent pathways.

Journal of biological regulators and homeostatic agents·2015
Same journal

Beyond strain-specific immunity: Conserved antigenic targets, emerging platforms, and translational challenges in universal influenza and pan-coronavirus vaccine development.

Journal of virological methods·2026
Same journal

Analytical characteristics of the NeuMoDx™ SARS-CoV-2 assay and clinical agreement with the BD MAX system.

Journal of virological methods·2026
Same journal

Development of a real-time PCR assay for tracking the spatiotemporal dynamics of a cyanophage and its Synechococcus host in waters of shrimp aquaculture ecosystems.

Journal of virological methods·2026
Same journal

A universal RdRp-targeted primer set for broad, non-specific detection of Quinvirinae and other Betaflexiviridae infecting stone fruits.

Journal of virological methods·2026
Same journal

Development of a multiplex real-time PCR method for detecting immunosuppressive viruses and its preliminary application in broilers.

Journal of virological methods·2026
Same journal

Development of a quantum dot-based immunochromatographic test strip for rapid detection of infectious bursal disease virus.

Journal of virological methods·2026
See all related articles

This study introduces a new assay for reverse transcriptase (RT) activity using non-radioactive digoxigenin-labelled nucleotides. The method offers a sensitive, specific, and easy-to-perform alternative for quantifying retroviral enzyme activity.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Virology

Background:

  • Reverse transcriptase (RT) is a key enzyme in retroviral replication.
  • Traditional assays for RT activity often rely on radioactive nucleotides, posing handling and disposal challenges.
  • Existing methods for measuring enzyme activity can be complex and difficult to standardize.

Purpose of the Study:

  • To develop a novel, sensitive, and non-radioactive assay for quantifying reverse transcriptase (RT) activity.
  • To improve upon existing methods by simplifying the detection process and enhancing standardization.
  • To provide a more accessible and user-friendly alternative for researchers studying retroviruses.

Main Methods:

  • Utilized digoxigenin-labelled dUTP instead of radioactive nucleotides for DNA synthesis.

Related Experiment Videos

  • Incorporated low concentrations of biotin-labelled dUTP to facilitate DNA immobilization.
  • Employed streptavidin-coated ELISA wells for capturing newly synthesized DNA.
  • Quantified enzyme activity via photometric detection using anti-digoxigenin antibodies and a colorimetric substrate (ABTSR).

    • Main Results:

    • The developed assay demonstrated high sensitivity and specificity for detecting RT activity.
    • The method successfully avoids the need for separating unincorporated nucleotides from synthesized DNA.
    • Standardization of RT activity measurement in enzyme units (nmol labelled dNTP incorporated/10 min at 37°C) is proposed over cpm.
    • The assay is described as easy to perform, making it suitable for routine laboratory use.

    Conclusions:

    • A novel, non-radioactive, and sensitive assay for reverse transcriptase activity has been successfully developed.
    • This assay offers significant advantages in terms of simplicity, standardization, and safety compared to traditional radioactive methods.
    • The proposed unit system for reporting RT activity enhances comparability across different studies and laboratories.