New and rapid visual detection assay for Trogoderma granarium everts based on recombinase polymerase amplification and CRISPR/Cas12a
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027006%3A_____%2F23%3A10176777" target="_blank" >RIV/00027006:_____/23:10176777 - isvavai.cz</a>
Alternative codes found
RIV/60460709:41210/23:95470
Result on the web
<a href="https://onlinelibrary.wiley.com/toc/15264998/2023/79/12" target="_blank" >https://onlinelibrary.wiley.com/toc/15264998/2023/79/12</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/ps.7739" target="_blank" >10.1002/ps.7739</a>
Alternative languages
Result language
angličtina
Original language name
New and rapid visual detection assay for Trogoderma granarium everts based on recombinase polymerase amplification and CRISPR/Cas12a
Original language description
BACKGROUND: Khapra beetle (Trogoderma granarium Everts), one of the most important quarantine pests globally, is capable of causing severe infestation and huge economic loss to stored grain, and its interception rate has increased in major global trade countries over the past few years. However, difficulties remain in distinguishing this species with similar ones. In order to assist border ports and warehouses in khapra beetle's effective rapid identification as well as pest control at the early stages of monitoring or interception, we herein developed a new and rapid visual detection assay for T. granarium based on recombinase polymerase amplification (RPA) and the CRISPR/Cas12a system. RESULTS: We designed and selected the first khapra beetle-specific RPA primers and crRNA, and optimized the visualization reaction system (Cas12a/CrRNA = 100 nM/500 nM). With only a 37 oC-heat-source and a blue light torch, RPA and CRISPR/CAS12a-based visualization assays can be completed within 40 min to differentiate between khapra beetle and nine similar Dermestidae species. After DNA extraction using a kit (4-5 h) or a simple method (5 min), the specific amplicons were obtained after a 15 min RPA reaction at 37 oC, followed by a 15 min color reaction under 37 oC in dark conditions using a CRISPR/CAS12a system and a fluorescent probe (5'-FAM/3'-BHQ1 labeled). This method is ingenious to low levels of DNA (10MINUS SIGN 1 ng μLMINUS SIGN 1) and meets the sensitivity requirements for detecting a single khapra beetle's egg (ALMOST EQUAL TO0.7 mm). CONCLUSION: Our specificity and sensitivity analysis inferred that the present visualization system is effective to quickly and uniquely detect khapra beetle at room temperature (37 oC), thereby preventing this species before they spread widely. Our study is suitable for being pushed forward in storage pest management, and provides value as a reference for monitoring and identification of other pests.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
40106 - Agronomy, plant breeding and plant protection; (Agricultural biotechnology to be 4.4)
Result continuities
Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2023
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
PEST MANAGEMENT SCIENCE
ISSN
1526-498X
e-ISSN
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Volume of the periodical
79
Issue of the periodical within the volume
12
Country of publishing house
GB - UNITED KINGDOM
Number of pages
8
Pages from-to
"5304 "- 5311
UT code for WoS article
001062914000001
EID of the result in the Scopus database
2-s2.0-85170521486