Preliminary results of the recombinase polymerase amplification technique for the detection of Haemonchus contortus from Hungarian field samples

Khangembam, R., Vass, N., Morrison, A., Melville, L. A., Antonopoulos, A. and Czeglédi, L. (2023) Preliminary results of the recombinase polymerase amplification technique for the detection of Haemonchus contortus from Hungarian field samples. Veterinary Parasitology, 320, 109974. (doi: 10.1016/j.vetpar.2023.109974) (PMID:37354888)

Abstract

Haemonchus contortus is a parasitic nematode of small ruminants responsible for significant economic losses and animal health concerns globally. Detection of gastrointestinal nematode (GIN) infection in veterinary practice typically relies on microscopy-based methods such as the faecal egg count and morphological identification of larval culture. However, mixed co-infections are common and species-specific identification is typically time-consuming and expertise-intensive. Compounded by increasing anthelmintic resistance, there is an urgent need to implement the molecular diagnosis of GIN in the livestock industry, preferably in field settings. Advances in isothermal amplification techniques including recombinase polymerase amplification (RPA) assays could improve this. Yet, constraints in RPA kit availability and amplicon detection systems limit the use of this technology in point of care settings. In this study, we present an early-stage, proof-of-concept demonstration of RPA targeting the internal transcribed spacer (ITS2) region of H. contortus. Having tested against eight closely related nematodes and also against five farm isolates in Eastern Hungary, preliminary results derived from a comparative analysis of 3 primer sets showed the assay detects H. contortus DNA and has a limit of detection of 10 ng/μl. We also tested an end-result naked eye detection system using various DNA binding dyes, of which EvaGreen® dye was successful for a qualitative RPA detection that could be adaptable at farm sites. [Abstract copyright: Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.]

• Item Type: Articles
• Additional Information: The study was supported by the EFOP-3.6.3-VEKOP-16-2017-00008 project which is co-financed by the European Union and the European Social Fund.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Antonopoulos, Mr Alistair

Creator Roles

Antonopoulos, A.: Conceptualization, Formal analysis, Methodology, Writing – review and editing, Writing – original draft

• Authors: Khangembam, R., Vass, N., Morrison, A., Melville, L. A., Antonopoulos, A., and Czeglédi, L.
• College/School: College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Journal Name: Veterinary Parasitology
• Publisher: Elsevier
• ISSN: 0304-4017
• ISSN (Online): 1873-2550
• Published Online: 22 June 2023
• Copyright Holders: Copyright © TheAuthor(s) 2023
• First Published: First published in Veterinary Parasitology 320:109974
• Publisher Policy: Reproduced under a creative commons licence