Establishment of a rapid method for skipjack tuna (Katsuwonus pelamis) authentication using molecular beacons in loop-mediated isothermal amplification. (15th July 2022)

• Record Type: Journal Article
• Authors: Xu, Wenjie; Li, Qiuping; Xue, Hanyue; Fei, Yanjin; Cui, Xiaowen; Cao, Min; Xiong, Xiaohui; Xiong, Xiong; Yang, Ying; Wang, Libin
• Is Part Of: Food chemistry. Volume 382(2022)
• Publication Date: 2022-07-15
• Subjects: molecular beacon (MB) -- loop-mediated isothermal amplification (LAMP) -- Skipjack tuna -- Species identification -- Target-specific detection; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2022.132365
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: Molecular beacon for Katsuwonus pelamis was designed and optimized. The real-time MB-LAMP assay was highly specific to Katsuwonus pelamis . Nonspecific amplification was not detected using the real-time MB-LAMP assay. Katsuwonus pelamis was detected from only 6 products (6/60). Abstract: One major drawback to the traditional loop-mediated isothermal amplification (LAMP) detection methods is the increased likelihood of detecting false-positive signals derived from non-specific amplification. Molecular beacon (MB) is increasingly being used in many applications and the MB-LAMP assay has proved itself as a target-specific method. The present work selected skipjack tuna as a case study, and developed a novel MB-LAMP assay for rapid species authentication. Specifically, the optimal MB structure includes 13 nucleobases in the loop region (binding specifically to loop primer LF) and 5 nucleobases in the stem region. For the established MB-LAMP assay, in the presence of the amplicons, the MB probe LFP-1 hybridizes to its target and forms a double helix. The change in conformation separates the quencher from the fluorophore, thereby resulting in the fluorescence release. The novel MB-LAMP assay has proved its specificity and can detect as little as 0.5 pg of skipjack tuna DNA.