Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples
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 thi...
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EDP Sciences
2021-01-01
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| Series: | Parasite |
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| Online Access: | https://www.parasite-journal.org/articles/parasite/full_html/2021/01/parasite210006/parasite210006.html |
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| author | Khangembam Rojesh Tóth Mariann Vass Nóra Várady Marián Czeglédi Levente Farkas Róbert Antonopoulos Alistair |
| author_facet | Khangembam Rojesh Tóth Mariann Vass Nóra Várady Marián Czeglédi Levente Farkas Róbert Antonopoulos Alistair |
| author_sort | Khangembam Rojesh |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T14:03:59Z |
| format | Article |
| id | doaj.art-2484493af91f486a956972068d05a762 |
| institution | Directory Open Access Journal |
| issn | 1776-1042 |
| language | English |
| last_indexed | 2024-03-11T14:03:59Z |
| publishDate | 2021-01-01 |
| publisher | EDP Sciences |
| record_format | Article |
| series | Parasite |
| spelling | doaj.art-2484493af91f486a956972068d05a7622023-11-02T03:00:34ZengEDP SciencesParasite1776-10422021-01-01288210.1051/parasite/2021078parasite210006Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samplesKhangembam RojeshTóth MariannVass Nóra0Várady Marián1https://orcid.org/0000-0001-6589-8219Czeglédi Levente2Farkas Róbert3Antonopoulos Alistair4Department of Animal Science, Institute of Animal Science, Biotechnology and Nature Conservation, Faculty of Agricultural and Food Sciences and Environmental Management, Böszörményi ut. 138, University of DebrecenInstitute of Parasitology, Slovak Academy of Sciences, Hlinkova 3Department of Animal Science, Institute of Animal Science, Biotechnology and Nature Conservation, Faculty of Agricultural and Food Sciences and Environmental Management, Böszörményi ut. 138, University of DebrecenDepartment of Parasitology and Zoology, University of Veterinary MedicineSchool of Veterinary Medicine, College of Medical, Veterinary and Life Sciences, Garscube Campus Bearsden Road, University of GlasgowIn 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.https://www.parasite-journal.org/articles/parasite/full_html/2021/01/parasite210006/parasite210006.htmllamp assaylateral flowhaemonchus contortuspoint-of-care diagnosisfill-flotac® |
| spellingShingle | Khangembam Rojesh Tóth Mariann Vass Nóra Várady Marián Czeglédi Levente Farkas Róbert Antonopoulos Alistair Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples Parasite lamp assay lateral flow haemonchus contortus point-of-care diagnosis fill-flotac® |
| title | Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples |
| title_full | Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples |
| title_fullStr | Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples |
| title_full_unstemmed | Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples |
| title_short | Point of care colourimetric and lateral flow LAMP assay for the detection of Haemonchus contortus in ruminant faecal samples |
| title_sort | point of care colourimetric and lateral flow lamp assay for the detection of haemonchus contortus in ruminant faecal samples |
| topic | lamp assay lateral flow haemonchus contortus point-of-care diagnosis fill-flotac® |
| url | https://www.parasite-journal.org/articles/parasite/full_html/2021/01/parasite210006/parasite210006.html |
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