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Use of Degenerate Primers in the Polymerase Chain Reaction to Detect Whitefly-Transmitted Geminiviruses
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1993
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GeneticsMolecular BiologyGenomicsPolymerase Chain ReactionDegenerate PrimersVirus PhylogenyMolecular DiagnosticsVirus GenePlant VirusDegenerate Pcr PrimersInsect VirusDna ReplicationVirologyWhitefly-transmitted GeminivirusesVirus ClassificationNatural SciencesPathogenesisPcr PrimersMedicineGenome Editing
Geminiviruses pose a serious threat to vegetable production in tropical and subtropical regions, necessitating accurate identification, and are amenable to PCR due to their double‑stranded circular DNA. Degenerate PCR primers targeting conserved sequences of 10 whitefly‑transmitted geminiviruses were designed and successfully amplified DNA‑A and/or DNA‑B fragments from 15 previously uncharacterized geminiviruses across the Americas, Caribbean Basin, and Africa.
Geminiviruses are widely recognised as a serious threat to vegetable production in many tropical and subtropical regions. This has increased the need for accurate identification of these viruses. Geminiviruses are well suited to polymerase chain reaction (PCR) methods because they replicate via a double-stranded, circular DNA form. Degenerate PCR primers were designed to anneal to highly conserved nucleotide sequences identified in the genomes of 10 whitefly-transmitted geminiviruses. The PCR primers were tested for their effectiveness in the amplification of viral DNA fragments from the DNA-A and/or DNA-B components of 15 previously uncharacterized geminiviruses from the Americas, the Caribbean Basin, and Africa (.)