Development and evaluation of a novel visual and rapid detection assay for toxigenic Fusarium graminearum in maize based on recombinase polymerase amplification and lateral flow analysis.
[en] Maize ear rot caused by Fusarium graminearum is one of the most severe maize diseases in global maize-growing regions. It reduces maize yield in the field and is also responsible for mycotoxin contamination of grains during the postharvest period. F. graminearum is one of the major deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEN) producers. The ingestion of these mycotoxins represents a risk for human and animal health. Hence, early detection and identification of F. graminearum are crucial to controlling these mycotoxins along the food or feed supply chains. In this study, the recombinase polymerase amplification with lateral flow dipstick (RPA-LFD) assay targeting the gaoA gene that codes for galactose oxidase was developed. The reaction conditions were optimized to make the method rapid, sensitive, and cost-effective. The developed RPA-LFD assay could detect the presence of 20 fg of the target genomic DNA per reaction within 25 min at 40 °C. Moreover, 52 field samples were tested using the developed RPA-LFD assay and compared with conventional PCR-based methods. The positive rate between RPA-LFD and the conventional PCR-based method was 100%. In conclusion, the developed method provides a novel alternative for the rapid, sensitive, and specific detection and identification of F. graminearum. It is not only workable for bulk maize samples without using sophisticated lab equipment but is also potentially useful for other agriculturally important toxigenic fungi.
Disciplines :
Microbiology
Author, co-author :
Liang, Xiaoyan ✱; Université de Liège - ULiège > TERRA Research Centre
Zhang, Xiu ✱; North Minzu University, Ningxia Key Laboratory for the Development and Application of Microbial Resources in Extreme Environments, Yinchuan 750021, Ningxia Hui Autonomous Region, PR China
Haseeb, Hafiz Abdul; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, PR China
Tang, Tingting; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, PR China
Shan, Jihao; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, PR China. Electronic address: shanjihao2007@163.com
Yin, Bo; National University of Singapore (Suzhou) Research Institute, Suzhou, Jiangsu, PR China
Guo, Wei; Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Beijing 100193, PR China. Electronic address: guowei01@caas.cn
✱ These authors have contributed equally to this work.
Language :
English
Title :
Development and evaluation of a novel visual and rapid detection assay for toxigenic Fusarium graminearum in maize based on recombinase polymerase amplification and lateral flow analysis.
Publication date :
02 July 2022
Journal title :
International Journal of Food Microbiology
ISSN :
0168-1605
eISSN :
1879-3460
Publisher :
Elsevier, Netherlands
Volume :
372
Pages :
109682
Peer reviewed :
Peer Reviewed verified by ORBi
Funding text :
This work was supported by the National Key Research and Development Program of China ( 2017YFC1600903 ), National Natural Science Foundation of China (No. 32072377 and 31670143 ), Beijing Natural Science Foundation (No. 6192023 ), Agricultural Science and Technology Innovation Program ( CAAS-ASTIP-2020-IFST-03 ), and The Open Foundation of Ningxia Key Laboratory for the Development and Application of Microbial Resources in Extreme Environments ( 2019TSWZ02 ).
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