Antonopoulos, A., Doyle, S. R., Bartley, D. J., Morrison, A. A., Kaplan, R., Howell, S., Neveu, C., Busin, V. , Devaney, E. and Laing, R. (2022) Allele specific PCR for a major marker of levamisole resistance in Haemonchus contortus. International Journal for Parasitology: Drugs and Drug Resistance, 20, pp. 17-26. (doi: 10.1016/j.ijpddr.2022.08.001) (PMID:35970104) (PMCID:PMC9399269)
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Abstract
Haemonchus contortus is a haematophagous parasitic nematode that infects small ruminants and causes significant animal health concerns and economic losses within the livestock industry on a global scale. Treatment primarily depends on broad-spectrum anthelmintics, however, resistance is established or rapidly emerging against all major drug classes. Levamisole (LEV) remains an important treatment option for parasite control, as resistance to LEV is less prevalent than to members of other major classes of anthelmintics. LEV is an acetylcholine receptor (AChR) agonist that, when bound, results in paralysis of the worm. Numerous studies implicated the AChR sub-unit, ACR-8, in LEV sensitivity and in particular, the presence of a truncated acr-8 transcript or a deletion in the acr-8 locus in some resistant isolates. Recently, a single non-synonymous SNP in acr-8 conferring a serine-to-threonine substitution (S168T) was identified that was strongly associated with LEV resistance. Here, we investigate the role of genetic variation at the acr-8 locus in a controlled genetic cross between the LEV susceptible MHco3(ISE) and LEV resistant MHco18(UGA2004) isolates of H. contortus. Using single worm PCR assays, we found that the presence of S168T was strongly associated with LEV resistance in the parental isolates and F3 progeny of the genetic cross surviving LEV treatment. We developed and optimised an allele-specific PCR assay for the detection of S168T and validated the assay using laboratory isolates and field samples that were phenotyped for LEV resistance. In the LEV-resistant field population, a high proportion (>75%) of L3 encoded the S168T variant, whereas the variant was absent in the susceptible isolates studied. These data further support the potential role of acr-8 S168T in LEV resistance, with the allele-specific PCR providing an important step towards establishing a sensitive molecular diagnostic test for LEV resistance.
| 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]. SRD is supported by a UKRI Future Leaders Fellowship [MR/T020733/1] and RL is supported by a Wellcome Clinical Research Career Development Fellowship [216614/Z/19/Z]. DB and AM are supported by the Scottish Government's Rural and Environment Science and Analytical Services (RESAS) division. |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Devaney, Professor Eileen and Busin, Valentina and Antonopoulos, Mr Alistair and Laing, Dr Roz |
| Authors: | Antonopoulos, A., Doyle, S. R., Bartley, D. J., Morrison, A. A., Kaplan, R., Howell, S., Neveu, C., Busin, V., Devaney, E., and Laing, R. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine |
| Journal Name: | International Journal for Parasitology: Drugs and Drug Resistance |
| Publisher: | Elsevier |
| ISSN: | 2211-3207 |
| ISSN (Online): | 2211-3207 |
| Published Online: | 10 August 2022 |
| Copyright Holders: | Copyright © 2022 The Authors |
| First Published: | First published in International Journal for Parasitology: Drugs and Drug Resistance 20: 17-26 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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