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Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces

Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targe...

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Autores principales: Wu, Yao-Dong, Wang, Qi-Qi, Wang, Meng, Elsheikha, Hany M., Yang, Xin, Hu, Min, Zhu, Xing-Quan, Xu, Min-Jun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Society for Parasitology and Tropical Medicine 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8106983/
https://www.ncbi.nlm.nih.gov/pubmed/33951773
http://dx.doi.org/10.3347/kjp.2021.59.2.167
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author Wu, Yao-Dong
Wang, Qi-Qi
Wang, Meng
Elsheikha, Hany M.
Yang, Xin
Hu, Min
Zhu, Xing-Quan
Xu, Min-Jun
author_facet Wu, Yao-Dong
Wang, Qi-Qi
Wang, Meng
Elsheikha, Hany M.
Yang, Xin
Hu, Min
Zhu, Xing-Quan
Xu, Min-Jun
author_sort Wu, Yao-Dong
collection PubMed
description Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation.
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spelling pubmed-81069832021-05-17 Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces Wu, Yao-Dong Wang, Qi-Qi Wang, Meng Elsheikha, Hany M. Yang, Xin Hu, Min Zhu, Xing-Quan Xu, Min-Jun Korean J Parasitol Brief Communication Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of Haemonchus contortus in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected H. contortus DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of H. contortus and merits further evaluation. The Korean Society for Parasitology and Tropical Medicine 2021-04 2021-04-30 /pmc/articles/PMC8106983/ /pubmed/33951773 http://dx.doi.org/10.3347/kjp.2021.59.2.167 Text en © 2021, Korean Society for Parasitology and Tropical Medicine https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0 (https://creativecommons.org/licenses/by-nc/4.0/) ) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Brief Communication
Wu, Yao-Dong
Wang, Qi-Qi
Wang, Meng
Elsheikha, Hany M.
Yang, Xin
Hu, Min
Zhu, Xing-Quan
Xu, Min-Jun
Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces
title Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces
title_full Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces
title_fullStr Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces
title_full_unstemmed Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces
title_short Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces
title_sort development of a lateral flow strip-based recombinase polymerase amplification assay for the detection of haemonchus contortus in goat feces
topic Brief Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8106983/
https://www.ncbi.nlm.nih.gov/pubmed/33951773
http://dx.doi.org/10.3347/kjp.2021.59.2.167
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