2026-04-14T04:00:55Zhttps://recercat.cat/oai/requestoai:recercat.cat:20.500.12327/29352025-10-22T11:06:26Zcom_2072_4428com_2072_4427col_2072_487898
RECERCAT
author
Bohórquez, Jose Alejandro
author
Muñoz-Aguilera, Adriana
author
Lanka, Saraswathi
author
Coronado, Liani
author
Rosell, Rosa
author
Alberch, Mònica
author
Maddox, Carol W.
author
Ganges, Llilianne
other
Producció Animal
group
Sanitat Animal
2025-10-22T11:06:26Z
2025-10-22T11:06:26Z
2024-04-15
Bohórquez, JA, Muñoz-Aguilera A, Lanka S, Coronado L, Rosell R, Alberch M, Maddox CW and Ganges L. 2024. “Development of a new loop-mediated isothermal amplification test for the sensitive, rapid, and economic detection of different genotypes of Classical swine fever virus”. Frontiers in Cellular and Infection Microbiology 14: 1372166. doi: 10.3389/fcimb.2024.1372166
2235-2988
http://hdl.handle.net/20.500.12327/2935
https://doi.org/10.3389/fcimb.2024.1372166
Background: Classical swine fever virus (CSFV) remains one of the most
important pathogens in animal health. Pathogen detection relies on viral RNA
extraction followed by RT-qPCR. Novel technologies are required to improve
diagnosis at the point of care.
Methods: A loop-mediated isothermal amplification (LAMP) PCR technique was
developed, with primers designed considering all reported CSFV genotypes. The
reaction was tested using both fluorometric and colorimetric detection, in
comparison to the gold standard technique. Viral strains from three circulating
CSFV genotypes were tested, as well as samples from infected animals. Other
pathogens were also tested, to determine the LAMP specificity. Besides
laboratory RNA extraction methods, a heating method for RNA release, readily
available for adaptation to field conditions was evaluated.
Results: Three primer sets were generated, with one of them showing better
performance. This primer set proved capable of maintaining optimal
performance at a wide range of amplification temperatures (60°C - 68°C). It
was also able to detect CSFV RNA from the three genotypes tested. The assay
was highly efficient in detection of samples from animals infected with field
strains from two different genotypes, with multiple matrices being detected using
both colorimetric and fluorometric methods. The LAMP assay was negative for all
the unrelated pathogens tested, including Pestiviruses. The only doubtful result in
both fluorometric and colorimetric LAMP was against the novel Pestivirus
italiaense, ovine Italy Pestivirus (OVPV), which has proven to have crossreaction with multiple CSFV diagnostic techniques. However, it is only possible to
detect the OVPV in a doubtful result if the viral load is higher than 10000
viral particles.
Conclusion: The results from the present study show that LAMP could be an
important addition to the currently used molecular diagnostic techniques for
CSFV. This technique could be used in remote locations, given that it can be
adapted for successful use with minimal equipment and minimally invasive
samples. The joined use of novel and traditional diagnostic techniques could
prove to be a useful alternative to support the CSF control.
eng
Attribution 4.0 International
Development of a new loop-mediated isothermal amplification test for the sensitive, rapid, and economic detection of different genotypes of Classical swine fever virus
info:eu-repo/semantics/article