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Real-Time PCR Protocol for Phytoplasma Detection and Quantification

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Phytoplasmas

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1875))

Abstract

Phytoplasmas are mollicutes restricted to plant phloem tissue and are normally present at very low concentrations. Real-time polymerase chain reaction (qPCR) offers several advantages over conventional PCR. It is a fast, sensitive, and reliable detection technique amenable to high throughput. Two fluorescent chemistries are available, intercalating dyes or hybridization probes. Intercalating dyes are relatively less expensive than TaqMan® hybridization probes but the latter chemistry is the most commonly used for phytoplasma detection. qPCR may be designed for universal detection of phytoplasma, group or subgroup specific detection, or for simultaneous detection of up to three or four phytoplasmas (multiplexing). qPCR may be used for relative or absolute quantification in host plants and in insect vectors. Therefore, qPCR plays an important role in phytoplasma detection as well as in host-pathogen interaction and in epidemiological studies. This chapter outlines the protocols followed in qPCR assay for phytoplasma detection and quantification, focusing mainly on the use of TaqMan® probes.

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Author information

Authors and Affiliations

  1. Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon

    Yusuf Abou-Jawdah, Vicken Aknadibossian, Maan Jawhari & Patil Tawidian

  2. Gulf Coast and Research Education Center, Wimauma, FL, USA

    Peter Abrahamian

Authors
  1. Yusuf Abou-Jawdah
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  2. Vicken Aknadibossian
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  3. Maan Jawhari
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  4. Patil Tawidian
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  5. Peter Abrahamian
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Corresponding author

Correspondence to Yusuf Abou-Jawdah .

Editor information

Editors and Affiliations

  1. Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy

    Rita Musetti

  2. Department of Agricultural, Food, Environmental and Animal Sciences, University of Udine, Udine, Italy

    Laura Pagliari

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Abou-Jawdah, Y., Aknadibossian, V., Jawhari, M., Tawidian, P., Abrahamian, P. (2019). Real-Time PCR Protocol for Phytoplasma Detection and Quantification. In: Musetti, R., Pagliari, L. (eds) Phytoplasmas. Methods in Molecular Biology, vol 1875. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8837-2_9

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  • DOI: https://doi.org/10.1007/978-1-4939-8837-2_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8836-5

  • Online ISBN: 978-1-4939-8837-2

  • eBook Packages: Springer Protocols

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