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Molecular and Biological Characterization of a Newly Identified Virus Representing a Novel Taxon of <i>Alphaflexiviridae</i> Infecting Different Accessions of Seashore Paspalum, a Turfgrass, Widely Grown in the United States.

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Real-Time PCR Protocol for Phytoplasma Detection and Quantification.

Yusuf Abou-Jawdah1, Vicken Aknadibossian2, Maan Jawhari2

  • 1Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon. abujawyf@aub.edu.lb.

Methods in Molecular Biology (Clifton, N.J.)
|October 27, 2018
PubMed
Summary

Real-time PCR (qPCR) provides a fast, sensitive method for detecting phytoplasmas, which are plant pathogens. This technique, particularly using TaqMan® probes, aids in understanding plant-insect interactions and disease spread.

Keywords:
DiagnosisInternal control genesMollicutesPhytoplasmaQuantitationSYBR greenTaqMan® probesqPCR

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Area of Science:

  • Plant pathology
  • Molecular biology
  • Microbiology

Background:

  • Phytoplasmas are plant-pathogenic bacteria lacking cell walls, residing in phloem tissue at low concentrations.
  • Conventional detection methods for phytoplasmas are often slow and lack sensitivity.
  • Real-time polymerase chain reaction (qPCR) has emerged as a superior alternative for pathogen detection.

Purpose of the Study:

  • To outline protocols for phytoplasma detection and quantification using qPCR.
  • To highlight the advantages of qPCR over conventional PCR for phytoplasma analysis.
  • To focus on the application of TaqMan® probes in qPCR assays for phytoplasma research.

Main Methods:

  • Utilizing real-time polymerase chain reaction (qPCR) for sensitive and rapid phytoplasma detection.
  • Employing fluorescent chemistries, specifically TaqMan® hybridization probes, for enhanced specificity.
  • Designing qPCR assays for universal, group-specific, or multiplexed phytoplasma detection.

Main Results:

  • qPCR demonstrates high sensitivity, speed, and reliability for detecting low-concentration phytoplasmas.
  • TaqMan® probes offer superior specificity and are widely adopted for phytoplasma detection compared to intercalating dyes.
  • qPCR enables both relative and absolute quantification of phytoplasmas in plants and insect vectors.

Conclusions:

  • qPCR is a crucial tool for accurate phytoplasma detection and quantification.
  • The technique significantly contributes to host-pathogen interaction studies and epidemiological investigations.
  • Standardized qPCR protocols, especially with TaqMan® probes, are essential for reliable phytoplasma research.