Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique
Abstract 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‐...
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| Format: | Article |
| Language: | English |
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Wiley
2021-07-01
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| Series: | Veterinary Medicine and Science |
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| Online Access: | https://doi.org/10.1002/vms3.455 |
| _version_ | 1818907578755710976 |
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| author | Siamak Amiri Bahar Shemshadi Saloomeh Shirali Farnaz Kheirandish Shirzad Fallahi |
| author_facet | Siamak Amiri Bahar Shemshadi Saloomeh Shirali Farnaz Kheirandish Shirzad Fallahi |
| author_sort | Siamak Amiri |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-19T21:57:21Z |
| format | Article |
| id | doaj.art-0629557544574ae38b166188e5654efe |
| institution | Directory Open Access Journal |
| issn | 2053-1095 |
| language | English |
| last_indexed | 2024-12-19T21:57:21Z |
| publishDate | 2021-07-01 |
| publisher | Wiley |
| record_format | Article |
| series | Veterinary Medicine and Science |
| spelling | doaj.art-0629557544574ae38b166188e5654efe2022-12-21T20:04:14ZengWileyVeterinary Medicine and Science2053-10952021-07-01741316132410.1002/vms3.455Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) techniqueSiamak Amiri0Bahar Shemshadi1Saloomeh Shirali2Farnaz Kheirandish3Shirzad Fallahi4Department of Pathobiology, Science and Research branch Islamic Azad University Tehran IranDepartment of Pathobiology, Science and Research branch Islamic Azad University Tehran IranDepartment of Pathobiology, Science and Research branch Islamic Azad University Tehran IranDepartment of Parasitology and Mycology School of Medicine Lorestan University of Medical Sciences Khorramabad IranDepartment of Parasitology and Mycology School of Medicine Lorestan University of Medical Sciences Khorramabad IranAbstract 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.https://doi.org/10.1002/vms3.455accuratecopro‐DNAFasciola hepaticaLAMPrapid detectionsheep |
| spellingShingle | Siamak Amiri Bahar Shemshadi Saloomeh Shirali Farnaz Kheirandish Shirzad Fallahi Accurate and rapid detection of Fasciola hepatica copro‐DNA in sheep using loop‐mediated isothermal amplification (LAMP) technique Veterinary Medicine and Science accurate copro‐DNA Fasciola hepatica LAMP rapid detection sheep |
| 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 | accurate copro‐DNA Fasciola hepatica LAMP rapid detection sheep |
| url | https://doi.org/10.1002/vms3.455 |
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