期刊
FOOD ANALYTICAL METHODS
卷 15, 期 3, 页码 658-665出版社
SPRINGER
DOI: 10.1007/s12161-021-02159-1
关键词
Self-quenching probe; Loop-mediated isothermal amplification (LAMP); Skipjack tuna; Species identification; Target-specific detection
资金
- National Key Research and Development Program of China [2018YFC1602800]
- National Natural Science Foundation of China [31701688]
A novel LAMP assay coupled with a self-quenching fluorogenic probe was developed for rapid detection of skipjack tuna, achieving high sensitivity and tolerance, and successfully authenticating skipjack tuna from commercial tuna products.
Target-specific detection of LAMP amplicons can be achieved through the use of target-specific probes or modified primers as biorecognition elements, decreasing the likelihood of detecting false-positive signals. The present work selected skip-jack tuna as a case study, and developed a novel LAMP assay coupled with the self-quenching fluorogenic probe for rapid detection of skipjack tuna. Specifically, the inner forward primer (FIP) was identified as the candidate primer to design the self-quenching fluorogenic probe. The fluorophore attached to the FIP (FIP-FAM) is self-quenched in unbound state. After binding to the dumbbell-shaped DNA target specifically, the FAM fluorophore is de-quenched, resulting in the fluorescence development. With the novel assay, as little as 5 fg of skipjack tuna, DNA could be detected. Moreover, the novel assay embraces a higher tolerance to impurities than non-specific fluorescent dyes, and succeeded the authentication of skipjack tuna from 6 commercial tuna products.
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