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Rapid detection of sweepoviruses through lateral flow dipstick-based recombinase polymerase amplification.
Acta Virol ; 66(2): 186-191, 2022.
Article in En | MEDLINE | ID: mdl-35766476
ABSTRACT
Sweepoviruses represent a phylogenetic group of begomoviruses that cause significant sweet potato (Ipomoea batatas) production losses in various countries across the world. For rapid identification of sweepoviruses, we developed a technique based on isothermal recombinase polymerase amplification in conjunction with lateral flow dipsticks (RPA-LFD). The optimum reaction conditions for the RPA were 20 min incubation at 37°C. The RPA-LFD specifically detected distinct sweepovirus species, with no other viruses infecting sweet potato causing a cross-reaction. The detection limit of the RPA-LFD was 1.0×104 copies of the target DNA molecule per reaction, and it exhibited a 10-fold greater sensitivity than the conventional PCR. Furthermore, when coupled with an alkaline polyethylene glycol-based crude genomic DNA extraction, the entire procedure was completed in 30 min without the use of any special instruments other than a water bath. Therefore, the RPA-LFD technique is a potential sweepovirus diagnostic tool that can be used in the field with fewer available resources. Keywords detection; sweepoviruses; recombinase polymerase amplification; lateral flow dipstick.
Subject(s)

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Nucleic Acid Amplification Techniques / Recombinases Type of study: Diagnostic_studies Language: En Journal: Acta Virol Year: 2022 Type: Article

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Nucleic Acid Amplification Techniques / Recombinases Type of study: Diagnostic_studies Language: En Journal: Acta Virol Year: 2022 Type: Article