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Rapid detection of viral, bacterial, fungal, and oomycete pathogens on tomato with microneedles, LAMP on a microfluidic chip, and smartphone device.
Shymanovich, Tatsiana; Saville, Amanda C; Paul, Rajesh; Wei, Qingshan; Ristaino, Jean Beagle.
Afiliação
  • Shymanovich T; North Carolina State University at Raleigh, Entomology and Plant Pathology, Raleigh, North Carolina, United States; tshymanovi@pa.gov.
  • Saville AC; North Carolina State University, Entomology and Plant Pathology, 2503 Thomas Hall, Raleigh, North Carolina, United States, 27695; acsavill@ncsu.edu.
  • Paul R; North Carolina State University at Raleigh, Chemical and Biomolecular Engineering, Raleigh, North Carolina, United States; rajeshpaulche07@gmail.com.
  • Wei Q; North Carolina State University, Chemical and Biomolecular Engineering, 911 Partners Way, Campus Box 7905, Raleigh, North Carolina, United States, 27695; qwei3@ncsu.edu.
  • Ristaino JB; North Carolina State University at Raleigh, Entomol. & Plant Pathology, Raleigh, North Carolina, United States; jbr@ncsu.edu.
Phytopathology ; 2024 Jun 03.
Article em En | MEDLINE | ID: mdl-38829831
ABSTRACT
Rapid detection of plant diseases before they escalate can improve disease control. Our team has developed rapid nucleic acid extraction methods with microneedles (MN) and combined these with LAMP assays for pathogen detection in the field. In this work, we developed LAMP assays for early blight (Alternaria linariae, A. alternata, and A. solani) and bacterial spot of tomato (Xanthomonas perforans) and validated these LAMP assays and two previously developed LAMP assays for tomato spotted wilt virus and late blight. Tomato plants were inoculated and disease severity was measured. Extractions were performed using MN and LAMP assays were run in tubes (with hydroxynaphthol blue) on a heat block or on a newly designed microfluidic slide chip on a heat block or a slide heater. Fluorescence on the microfluidic chip slides was visualized using EvaGreen and photographed on a smartphone. Plants inoculated with X. perforans or tomato spotted wilt virus tested positive prior to visible disease symptoms, while P. infestans and A. linariae were detected at the time of visual disease symptoms. LAMP assays were more sensitive than PCR and the limit of detection was 1 pg of DNA for both A. linariae and X. perforans. The LAMP assay designed for early blight detected all three species of Alternaria that infect tomato and is thus an Alternaria spp. assay. This study demonstrates the utility of rapid MN extraction followed by LAMP on a microfluidic chip for rapid diagnosis of four important tomato pathogens.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Ano de publicação: 2024 Tipo de documento: Article