Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples

• Published in: Parasite (Paris) Vol. 28; p. 82
• Main Authors: Khangembam, Rojesh, Tóth, Mariann, Vass, Nóra, Várady, Marián, Czeglédi, Levente, Farkas, Róbert, Antonopoulos, Alistair
• Format: Journal Article
• Language: English
• Published: France EDP Sciences 2021
• Subjects: Animal sciences; Animals; Assaying; Centrifugation; Colorimetry; Deoxyribonucleic acid; Diagnostic systems; DNA; Eggs; Environmental management; Feces; fill-flotac; Fluctuations; Haemonchiasis - diagnosis; Haemonchiasis - veterinary; Haemonchus - genetics; Haemonchus contortus; Infections; lamp assay; lateral flow; Lysis; Microscopy; Molecular Diagnostic Techniques; Nucleic Acid Amplification Techniques; Parasites; Parasitology; point-of-care diagnosis; Point-of-Care Systems; Ruminants; Special Issue – Combatting Anthelmintic resistance in ruminants. Invited Editors: Johannes Charlier, Hervé Hoste, and Smaragda Sotiraki; Terrain; Veterinary medicine; Zoology; Point-of-Care Diagnosis; Haemonchus contortus; Fill-FLOTAC; LAMP assay; Lateral flow
• ISSN: 1776-1042; 1252-607X; 1776-1042
• DOI: 10.1051/parasite/2021078

In this study, we present an optimised colourimetric and a lateral flow LAMP assay for the detection of Haemonchus contortus in small ruminant faecal samples. Using a previously published LAMP primer set, we made use of commercially available colourimetric LAMP and lateral flow kits and combined this into an optimised diagnostic assay which was then tested on field faecal samples from Eastern and South-Eastern Hungary as well as a pure H. contortus egg faecal sample from Košice, Slovakia. Both assays showed no conflicts in visual detection of the results. Additionally, we modified and tested several centrifuge-free DNA extraction methods and one bead-beating egg lysis DNA extraction method to develop a true point of care protocol, as the source of the starting DNA is the main rate-limiting step in farm-level molecular diagnosis. Out of the various methods trialed, promising results were obtained with the magnetic bead extraction method. Sample solutions from the Fill-FLOTAC technique were also utilised, which demonstrated that it could be efficiently adapted for field-level egg concentration to extract DNA. This proof of concept study showed that isothermal amplification technologies with a colourimetric detection or when combined with a lateral flow assay could be an important step for a true point of care molecular diagnostic assay for H. contortus.