New and rapid visual detection assay for Trogoderma granarium everts based on recombinase polymerase amplification and CRISPR/Cas12a

Kód výsledku v 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>

Nalezeny alternativní kódy

RIV/60460709:41210/23:95470

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

New and rapid visual detection assay for Trogoderma granarium everts based on recombinase polymerase amplification and CRISPR/Cas12a

Popis výsledku v původním jazyce

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&apos;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&apos;-FAM/3&apos;-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&apos;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.

Název v anglickém jazyce

New and rapid visual detection assay for Trogoderma granarium everts based on recombinase polymerase amplification and CRISPR/Cas12a

Popis výsledku anglicky

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&apos;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&apos;-FAM/3&apos;-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&apos;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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

40106 - Agronomy, plant breeding and plant protection; (Agricultural biotechnology to be 4.4)

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2023

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

PEST MANAGEMENT SCIENCE

ISSN

1526-498X

e-ISSN

—

Svazek periodika

79

Číslo periodika v rámci svazku

12

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

8

Strana od-do

"5304 "- 5311

Kód UT WoS článku

001062914000001

EID výsledku v databázi Scopus

2-s2.0-85170521486