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Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique

Fascioliasis is a parasitic infection caused by Fasciola spp. in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validate methods for the diagnosis of fascioliasis in animals. This study aimed to compare the loop‐mediated...

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Autores principales: Amiri, Siamak, Shemshadi, Bahar, Shirali, Saloomeh, Kheirandish, Farnaz, Fallahi, Shirzad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8294381/
https://www.ncbi.nlm.nih.gov/pubmed/33624950
http://dx.doi.org/10.1002/vms3.455
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author Amiri, Siamak
Shemshadi, Bahar
Shirali, Saloomeh
Kheirandish, Farnaz
Fallahi, Shirzad
author_facet Amiri, Siamak
Shemshadi, Bahar
Shirali, Saloomeh
Kheirandish, Farnaz
Fallahi, Shirzad
author_sort Amiri, Siamak
collection PubMed
description Fascioliasis is a parasitic infection caused by Fasciola spp. in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validate methods for the diagnosis of fascioliasis in animals. This study aimed to compare the loop‐mediated isothermal amplification (LAMP) technique with PCR assay for the diagnosis of F. hepatica in sheep. In this cross‐sectional study, 195 stool samples were collected from sheep for 3 months in Lorestan province, West of Iran. Specimens’ parasitological examination was performed by using the direct wet mount and formalin‐ether concentration method. After DNA extraction from the samples, molecular analysis was done using PCR and LAMP techniques based on the Fasciola ribosomal intergenic spacer (IGS) sequence. Of 195 specimens of sheep, 11 specimens were identified as F. hepatica‐positive infection by using microscopic, PCR and LAMP assays. Kappa agreement test results showed that there was a significant agreement between the results of microscopic examination diagnostic tests, PCR and LAMP (Kappa = 0.51–0.72 and p < .001). According to the results of chi‐square comparisons between parasite prevalence applying different techniques and variables of age, sex breed, and type of drinking water, there was no significant relationship (p ≥ .05). However, most of the infected sheep with Fasciola were 3‐ to 4‐year‐old females, of the Lori breed and consumed tap water. In many endemic areas, successful prevention and treatment of fascioliasis in animals depend on rapid and accurate diagnosis. Based on the results of the Kappa agreement, the significant agreement among the results of the microscopic examination, PCR and LAMP indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in sheep. However, molecular methods, especially the LAMP technique, are suggested because of their higher sensitivity and reliability for the diagnosis of F. hepatica even under field conditions.
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spelling pubmed-82943812021-07-23 Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique Amiri, Siamak Shemshadi, Bahar Shirali, Saloomeh Kheirandish, Farnaz Fallahi, Shirzad Vet Med Sci Original Articles Fascioliasis is a parasitic infection caused by Fasciola spp. in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validate methods for the diagnosis of fascioliasis in animals. This study aimed to compare the loop‐mediated isothermal amplification (LAMP) technique with PCR assay for the diagnosis of F. hepatica in sheep. In this cross‐sectional study, 195 stool samples were collected from sheep for 3 months in Lorestan province, West of Iran. Specimens’ parasitological examination was performed by using the direct wet mount and formalin‐ether concentration method. After DNA extraction from the samples, molecular analysis was done using PCR and LAMP techniques based on the Fasciola ribosomal intergenic spacer (IGS) sequence. Of 195 specimens of sheep, 11 specimens were identified as F. hepatica‐positive infection by using microscopic, PCR and LAMP assays. Kappa agreement test results showed that there was a significant agreement between the results of microscopic examination diagnostic tests, PCR and LAMP (Kappa = 0.51–0.72 and p < .001). According to the results of chi‐square comparisons between parasite prevalence applying different techniques and variables of age, sex breed, and type of drinking water, there was no significant relationship (p ≥ .05). However, most of the infected sheep with Fasciola were 3‐ to 4‐year‐old females, of the Lori breed and consumed tap water. In many endemic areas, successful prevention and treatment of fascioliasis in animals depend on rapid and accurate diagnosis. Based on the results of the Kappa agreement, the significant agreement among the results of the microscopic examination, PCR and LAMP indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in sheep. However, molecular methods, especially the LAMP technique, are suggested because of their higher sensitivity and reliability for the diagnosis of F. hepatica even under field conditions. John Wiley and Sons Inc. 2021-02-24 /pmc/articles/PMC8294381/ /pubmed/33624950 http://dx.doi.org/10.1002/vms3.455 Text en © 2021 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Amiri, Siamak
Shemshadi, Bahar
Shirali, Saloomeh
Kheirandish, Farnaz
Fallahi, Shirzad
Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
title Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
title_full Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
title_fullStr Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
title_full_unstemmed Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
title_short Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
title_sort accurate and rapid detection of fasciola hepatica copro‐dna in sheep using loop‐mediated isothermal amplification (lamp) technique
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8294381/
https://www.ncbi.nlm.nih.gov/pubmed/33624950
http://dx.doi.org/10.1002/vms3.455
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