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Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs

Pentatrichomonas hominis is a trichomonad protozoan that infects the cecum and colon of humans and other mammals. It is a zoonotic pathogen that causes diarrhea in both animals and humans. As companion animals, dogs infected with P. hominis pose a risk of transmitting it to humans. Current methods,...

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Autores principales: Rong, Yao, Zhang, Xichen, Chen, Xuejiao, Li, Jianhua, Gong, Pengtao, Wang, Xiaocen, Li, Xin, Zhang, Xu, Yue, Taotao, Zhang, Hongbo, Zhou, Xiaofei, Zhang, Nan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10670101/
https://www.ncbi.nlm.nih.gov/pubmed/37998756
http://dx.doi.org/10.3390/cimb45110579
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author Rong, Yao
Zhang, Xichen
Chen, Xuejiao
Li, Jianhua
Gong, Pengtao
Wang, Xiaocen
Li, Xin
Zhang, Xu
Yue, Taotao
Zhang, Hongbo
Zhou, Xiaofei
Zhang, Nan
author_facet Rong, Yao
Zhang, Xichen
Chen, Xuejiao
Li, Jianhua
Gong, Pengtao
Wang, Xiaocen
Li, Xin
Zhang, Xu
Yue, Taotao
Zhang, Hongbo
Zhou, Xiaofei
Zhang, Nan
author_sort Rong, Yao
collection PubMed
description Pentatrichomonas hominis is a trichomonad protozoan that infects the cecum and colon of humans and other mammals. It is a zoonotic pathogen that causes diarrhea in both animals and humans. As companion animals, dogs infected with P. hominis pose a risk of transmitting it to humans. Current methods, such as direct smears and polymerase chain reaction (PCR), used for P. hominis detection have limitations, including low detection rates and the need for specialized equipment. Therefore, there is an urgent need to develop rapid, sensitive, and simple detection methods for clinical application. Recombinase polymerase amplification (RPA) has emerged as a technology for rapid pathogen detection. In this study, we developed a lateral flow dipstick (LFD)-RPA method based on the highly conserved SPO11-1 gene for detecting P. hominis infection by optimizing the primers, probes, and reaction conditions, and evaluating cross-reactivity with genomes of Giardia duodenalis and other parasites. The LFD-RPA method was then used to test 128 dog fecal samples collected from Changchun. The results confirmed the high specificity of the method with no cross-reactivity with the five other parasites. The lowest detection limit of the method was 10(2) copies/µL, and its sensitivity was 10(0) times higher than that of the conventional PCR method. Consistent with the positivity rate observed using nested PCR, 12 samples (out of 128) tested positive using this method (positivity rate, 9.38%). In conclusion, the LFD-RPA method developed in this study represents a simple and sensitive assay that allows for the rapid detection of P. hominis infection in dogs, especially in this field.
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spelling pubmed-106701012023-11-17 Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs Rong, Yao Zhang, Xichen Chen, Xuejiao Li, Jianhua Gong, Pengtao Wang, Xiaocen Li, Xin Zhang, Xu Yue, Taotao Zhang, Hongbo Zhou, Xiaofei Zhang, Nan Curr Issues Mol Biol Communication Pentatrichomonas hominis is a trichomonad protozoan that infects the cecum and colon of humans and other mammals. It is a zoonotic pathogen that causes diarrhea in both animals and humans. As companion animals, dogs infected with P. hominis pose a risk of transmitting it to humans. Current methods, such as direct smears and polymerase chain reaction (PCR), used for P. hominis detection have limitations, including low detection rates and the need for specialized equipment. Therefore, there is an urgent need to develop rapid, sensitive, and simple detection methods for clinical application. Recombinase polymerase amplification (RPA) has emerged as a technology for rapid pathogen detection. In this study, we developed a lateral flow dipstick (LFD)-RPA method based on the highly conserved SPO11-1 gene for detecting P. hominis infection by optimizing the primers, probes, and reaction conditions, and evaluating cross-reactivity with genomes of Giardia duodenalis and other parasites. The LFD-RPA method was then used to test 128 dog fecal samples collected from Changchun. The results confirmed the high specificity of the method with no cross-reactivity with the five other parasites. The lowest detection limit of the method was 10(2) copies/µL, and its sensitivity was 10(0) times higher than that of the conventional PCR method. Consistent with the positivity rate observed using nested PCR, 12 samples (out of 128) tested positive using this method (positivity rate, 9.38%). In conclusion, the LFD-RPA method developed in this study represents a simple and sensitive assay that allows for the rapid detection of P. hominis infection in dogs, especially in this field. MDPI 2023-11-17 /pmc/articles/PMC10670101/ /pubmed/37998756 http://dx.doi.org/10.3390/cimb45110579 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Rong, Yao
Zhang, Xichen
Chen, Xuejiao
Li, Jianhua
Gong, Pengtao
Wang, Xiaocen
Li, Xin
Zhang, Xu
Yue, Taotao
Zhang, Hongbo
Zhou, Xiaofei
Zhang, Nan
Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs
title Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs
title_full Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs
title_fullStr Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs
title_full_unstemmed Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs
title_short Development of an LFD-RPA Assay for Rapid Detection of Pentatrichomonas hominis Infection in Dogs
title_sort development of an lfd-rpa assay for rapid detection of pentatrichomonas hominis infection in dogs
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10670101/
https://www.ncbi.nlm.nih.gov/pubmed/37998756
http://dx.doi.org/10.3390/cimb45110579
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