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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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Subviral Agents01:29

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Subviral agents are infectious entities that resemble viruses but lack one or more viral components, such as a capsid or essential replication machinery. These agents include viroids, prions, and satellites, each possessing distinct structural and functional characteristics that influence their mode of infection and replication.Viroids are the simplest subviral agents, consisting of circular, single-stranded RNA molecules without a protein coat. They exclusively infect plants, relying entirely...
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Updated: Oct 11, 2025

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Detection of Viroids Using RT-qPCR.

Subhas Hajeri1, Georgios Vidalakis2, Raymond K Yokomi3

  • 1Citrus Pest Detection Program, Central California Tristeza Eradication Agency, Tulare, CA, USA. shajeri@cctea.org.

Methods in Molecular Biology (Clifton, N.J.)
|November 30, 2021
PubMed
Summary

Viroids, tiny infectious RNA pathogens, are detected using reverse transcription quantitative polymerase chain reaction (RT-qPCR). This method amplifies specific viroid RNA sequences for accurate crop disease diagnosis.

Keywords:
CEVdHSVdOne-step RT-qPCRRT-qPCRViroid

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Area of Science:

  • Plant Pathology
  • Molecular Biology
  • Virology

Background:

  • Viroids are the smallest known infectious agents, consisting of single-stranded, circular, naked RNA.
  • These pathogens cause significant diseases in economically important crops.
  • Traditional viroid detection methods have been enhanced by molecular techniques.

Purpose of the Study:

  • To describe the application of reverse transcription quantitative polymerase chain reaction (RT-qPCR) for viroid detection.
  • To highlight the efficiency and sensitivity of RT-qPCR in identifying viroid RNA.

Main Methods:

  • Viroid RNA is detected using sequence-specific primers.
  • Reverse transcriptase synthesizes a complementary DNA (cDNA) copy from the viroid RNA template.
  • RNase H removes the RNA template from the cDNA:RNA hybrid, enhancing PCR sensitivity.
  • The resulting cDNA is amplified via PCR to detect the viroid sequence.

Main Results:

  • RT-qPCR has transformed viroid detection methodologies.
  • The described method allows for sensitive amplification and detection of specific viroid sequences.
  • This technique is crucial for diagnosing viroid-induced crop diseases.

Conclusions:

  • RT-qPCR is a powerful and sensitive tool for detecting viroid pathogens.
  • Accurate viroid detection is essential for managing crop diseases and ensuring agricultural productivity.