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1.
Data Brief ; 20: 471-479, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30186897

RESUMO

Loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) are two rapid isothermal amplification methods for detecting three common fungal root pathogens of cool-season turfgrass: Gaeumannomyces avenae, Magnaporthiopsis poae and Ophiosphaerella korrae, "Detection of root-infecting fungi on cool-season turfgrasses using loop-mediated isothermal amplification and recombinase polymerase amplification" (Karakkat et al., 2018) [1]. The data provided here describe the information for designing primers and probes for LAMP and RPA, how specific they are for each of the three fungi, and the evaluation of RPA on field samples.

2.
J Microbiol Methods ; 151: 90-98, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29964073

RESUMO

Root-infecting fungal pathogens such as Gaeumannomyces avenae, Ophiosphaerella korrae, and Magnaporthiopsis poae cause extensive damage to amenity turfgrasses in temperate climates. The diseases they cause are difficult to diagnose by visual symptoms or microscopic inspection, and traditional polymerase chain reaction-based assays require large financial investments in equipment such as thermal cyclers and highly trained staff. The primary objective of this research was to develop fast and accurate detection assays for the three pathogens listed above that did not require the use of thermal cycling equipment. Loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) assays were developed for each pathogen based on known fungal cultures. The assays were tested on 27 samples received at the University of Wisconsin's Turfgrass Diagnostic Laboratory in 2016 and 2017 and both methods provided accurate diagnoses within about 30 min with minimal sample preparation. However, the RPA assays had lower levels of false positive contamination relative to the LAMP assays and are more likely to be effective in a field or diagnostic laboratory for improved turf root-pathogen detection.


Assuntos
Fungos/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Micoses/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Doenças das Plantas/microbiologia , Raízes de Plantas/microbiologia , Recombinases , Ascomicetos/genética , Ascomicetos/patogenicidade , Primers do DNA , Fungos/genética , Fungos/patogenicidade , Reação em Cadeia da Polimerase/métodos , Estações do Ano , Sensibilidade e Especificidade , Temperatura
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