RT Journal Article
T1 Evaluation of the effect of outer primer structure, and inner primer linker sequences, in the performance of Loop-mediated isothermal amplification
A1 Lamas Freire, Alexandre
A1 Azinheiro, Sarah
A1 Roumani, Foteini
A1 Prado Rodríguez, Marta
A1 Garrido Maestu, Alejandro
K1 Loop-mediated isothermal amplification
K1 Linker
K1 Dual-priming oligonucleotide
K1 DPO
K1 L. monocytogenes
AB Loop-mediated isothermal amplification, or LAMP, is nowadays the most popular isothermal nucleic acid amplification technique. This technique implements a minimum of four primers, named outer (F3/B3) and inner primers (FIP/BIP). The inner primers hybridize in two distinct regions, and some studies have reported that the usage of a linker, typically composed of four thymines, in the middle of these primers can improve assay performance. In addition to this, dual-priming oligonucleotides, DPO, have been reported to provide highly specific reducing non-specific amplifications. Considering the large number of primers implemented in LAMP assays, in the current study the suitability of DPO primers replacing regular outer primers; and their combination with different linker sequences in the inner primers were explored. The results demonstrated that replacing standard F3/B3 by DPO primers does not significantly affect that overall performance of the assay, and provides additional stability to temperature changes. This observations were consistent regardless the type of linker implemented in the inner primers, out of which in the current study a linker composed of thymines significantly outperformed the other options tested, most likely due to a combination of sequence and physical structure.
PB Elsevier
SN 1873-3573
YR 2023
FD 2023-05-04
LK https://hdl.handle.net/10347/43534
UL https://hdl.handle.net/10347/43534
LA eng
NO Talanta 260 (2023) 124642
NO Interreg Atlantic Area Program (EAPA_758/2018)
NO Fundação para a Ciência e a Tecnologia (SFRH/BD/140396/2018)
NO Fundação para a Ciência e Tecnologia through the Scientific Employment Stimulus Program (2021.02810.CEECIND)
DS Minerva
RD 28 abr 2026