Antonopoulos, A. et al. (2024) Real-time single base specific detection of the Haemonchus contortus S168T variant associated with levamisole resistance using loop-primer endonuclease cleavage loop mediated isothermal amplification. Molecular and Cellular Probes, 73, 101946. (doi: 10.1016/j.mcp.2023.101946) (PMID:38097144) (PMCID:PMC10884526)
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Abstract
Haemonchus contortus is a parasitic haematophagous nematode that primarily affects small ruminants and causes significant economic loss to the global livestock industry. Treatment of haemonchosis typically relies on broad-spectrum anthelmintics, resistance to which is an important cause of treatment failure. Resistance to levamisole remains less widespread than to other major anthelmintic classes, prompting the need for more effective and accurate surveillance to maintain its efficacy. Loop-primer endonuclease cleavage loop-mediated isothermal amplification (LEC-LAMP) is a recently developed diagnostic method that facilitates multiplex target detection with single nucleotide polymorphism (SNP) specificity and portable onsite testing. In this study, we designed a new LEC-LAMP assay and applied it to detect the levamisole resistance marker S168T in H. contortus. We explored multiplexing probes for both the resistant S168T and the susceptible S168 alleles in a single-tube assay. We then included a generic probe to detect the acr-8 gene in the multiplex assay, which could facilitate the quantification of both resistance markers and overall genetic material from H. contortus in a single step. Our results showed promising application of these technologies, demonstrating a proof-of-concept assay which is amenable to detection of resistance alleles within the parasite population, with the potential for multiplex detection, and point-of-care application enabled by lateral flow end-point detection. However, further optimisation and validation is necessary.
| Item Type: | Articles |
|---|---|
| Additional Information: | This work was funded by a James Herriot Scholarship (Glasgow University Vet Fund) and a Biotechnology and Biological Sciences Research Council (BBSRC) strategic Lola [BB/M003949]. MMS is supported by Egyptian Ministry of Higher Education [MM8/19]. JR is supported by a UK Medical Research Council grant (MR/V035401/1). SRD is supported by a UKRI Future Leaders Fellowship [MR/T020733/1] and the Wellcome Trust (UK) through core funding to the Wellcome Sanger Institute (UK) [206194]. RL is supported by a Wellcome Clinical Research Career Development Fellowship [216614/Z/19/Z] and a University of Glasgow Lord Kelvin Adam Smith Fellowship (LKAS). DB and AM are supported by the Scottish Government's Rural and Environment Science and Analytical Services (RESAS) division. This research was funded, in part, by the Wellcome Trust. |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Shalaby, Maha Mansour Mohamed and Devaney, Professor Eileen and Busin, Valentina and Reboud, Professor Julien and Antonopoulos, Mr Alistair and Laing, Dr Roz and Doyle, Dr Stephen |
| Creator Roles: | Antonopoulos, A.Conceptualization, Investigation, Methodology, Project administration, Resources, Software, Validation, Visualization, Writing – original draft, Writing – review and editing Doyle, S. R.Conceptualization, Investigation, Methodology, Writing – review and editing Shalaby, M. M.Conceptualization Shalaby, M. M. M.Investigation, Methodology, Resources, Validation, Visualization, Writing – review and editing Reboud, J.Investigation, Methodology, Resources, Writing – review and editing Devaney, E.Conceptualization, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Visualization, Writing – original draft, Writing – review and editing Laing, R.Conceptualization, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Visualization, Writing – original draft, Writing – review and editing Busin, V.Conceptualization, Funding acquisition, Investigation, Methodology, Project administration, Resources, Supervision, Visualization, Writing – original draft, Writing – review and editing |
| Authors: | Antonopoulos, A., Higgins, O., Doyle, S. R., Bartley, D., Morrison, A., Shalaby, M. M., Reboud, J., Devaney, E., Smith, T. J., Laing, R., and Busin, V. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine College of Science and Engineering > School of Engineering > Biomedical Engineering |
| Journal Name: | Molecular and Cellular Probes |
| Publisher: | Elsevier |
| ISSN: | 0890-8508 |
| ISSN (Online): | 1096-1194 |
| Published Online: | 19 December 2023 |
| Copyright Holders: | Copyright © 2023 The Authors |
| First Published: | First published in Molecular and Cellular Probes 73:101946 |
| Publisher Policy: | Reproduced under a Creative Commons license |
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