Journal
ACS NANO
Volume 13, Issue 6, Pages 6540-6549Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.9b00193
Keywords
plant disease; DNA extraction; microneedle patch; Phytophthora infestans; nucleic acid amplification; point-of-care diagnostics
Categories
Funding
- Chancellor's Faculty Excellence Program (Emerging Plant Disease and Global Food Security Cluster)
- Kenan Institute for Engineering, Technology & Science (KIETS) at NC State
- USDA iPIPE funds [2015-0097]
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In-field molecular diagnosis of plant diseases via nucleic acid amplification is currently limited by cumbersome protocols for extracting and isolating pathogenic DNA from plant tissues. To address this challenge, a rapid plant DNA extraction method was developed using a disposable polymeric microneedle (MN) patch. By applying MN patches on plant leaves, amplification-assay-ready DNA can be extracted within a minute from different plant species. MN-extracted DNA was used for direct polymerase chain reaction amplification of plant plastid DNA without purification. Furthermore, using this patch device, extraction of plant pathogen DNA (Phytophthora infestans) from both laboratory-inoculated and field-infected leaf samples was performed for detection of late blight disease in tomato. MN extraction achieved 100% detection rate of late blight infections for samples after 3 days of inoculation when compared to the conventional gold standard cetyltrimethylammonium bromide (CTAB)-based DNA extraction method and 100% detection rate for all blind field samples tested. This simple, cell-lysis-free, and purification-free DNA extraction method could be a transformative approach to facilitate rapid sample preparation for molecular diagnosis of various plant diseases directly in the field.
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