Abstract
Polymerase chain reaction (PCR) is the most widely used nucleo-based method for the detection of plant viruses. It is a primer-mediated in vitro reaction involving amplification of target nucleic acid sequences. A standard PCR is a three-step procedure: (1) denaturation at a high temperature (90–95 °C), (2) annealing of target specific primers (45–60 °C) and (3) primer extension by a thermostable DNA polymerase at 72 °C. Different variants of PCR have been developed ranging from conventional PCR to real-time PCR to improve the sensitivity and specificity for the detection of plant viruses. Some virus detection methods are: DNA-based PCR, nested PCR (nPCR), immunocapture PCR (IC-PCR), multiplex PCR (M-PCR), real-time PCR and DNA finger printing. Others are RNA-based reverse transcription (RT) PCR, real-time RT-PCR, IC-RT-PCR and AmpliDet RNA. All these methods enable a rapid and accurate detection and quantification of plant viruses. Protocols of PCR assays for the detection of plant viruses are discussed in this chapter.
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Bhat, A.I., Rao, G.P. (2020). Polymerase Chain Reaction. In: Characterization of Plant Viruses . Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0334-5_35
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DOI: https://doi.org/10.1007/978-1-0716-0334-5_35
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