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A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of toxigenic Fusarium temperatum in maize stalks and kernels

Shan, Liuying, Abdul Haseeb, Hafiz, Zhang, Jun, Zhang, Dandan, Jeffers, Daniel Paul, Dai, Xiaofeng, Guo, Wei
International journal of food microbiology 2019 v.291 pp. 72-78
Fusarium, corn, corn ears, corn stover, disease control, equipment, feeds, food chain, fumonisin B1, loop-mediated isothermal amplification, nucleotide sequences, pathogens, rapid methods, ribosomal DNA, risk assessment, screening, seeds, supply chain, toxigenic strains, China
Fusarium temperatum is an emerging maize pathogen that causes maize ear and stalk rot diseases and produces various mycotoxins including moniliformin, beauvericin, enniatins and fumonisin B1, which poses a potential risk to the human food or animal feed supply chains. Early detection of F. temperatum is crucial to prevent its derived mycotoxins from entering the food chain, and is also a useful tool in disease management practices. Here, we describe a loop-mediated isothermal amplification (LAMP) assay for rapid diagnosis of F. temperatum. The 28S ribosomal DNA sequences (28S rDNA) of F. temperatum were used to design a set of six primers. The reaction conditions were optimized for developing a fast assay with high specificity and sensitivity, and were able to detect the presence of less than 10 pg of target DNA per reaction within 60 min. Furthermore, the resulting amplicons were visualized by adding SYBR Green I to the reaction tubes. Suspected F. temperatum infected maize stalk samples collected from Yunnan province, China were identified using the developed LAMP assay. In conclusion, the method not only provides a rapid and specific screening for the existence of F. temperatum in a bulk of maize samples without using sophisticated equipment, but also is potentially useful for other agriculturally important toxigenic fungi.