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Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals

BACKGROUND: Schistosomiasis japonica is a common zoonosis. Domestic animals are the primary source of infection and play an important role in disease transmission. The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and, for this reason, diagnosti...

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Autores principales: Zhang, Xin, He, Chuan-Chuan, Liu, Jin-Ming, Li, Hao, Lu, Ke, Fu, Zhi-Qiang, Zhu, Chuan-Gang, Liu, Yi-Ping, Tong, Lai-Bao, Zhou, De-bao, Zha, Li, Hong, Yang, Jin, Ya-Mei, Lin, Jiao-Jiao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5390378/
https://www.ncbi.nlm.nih.gov/pubmed/28407808
http://dx.doi.org/10.1186/s40249-017-0298-y
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author Zhang, Xin
He, Chuan-Chuan
Liu, Jin-Ming
Li, Hao
Lu, Ke
Fu, Zhi-Qiang
Zhu, Chuan-Gang
Liu, Yi-Ping
Tong, Lai-Bao
Zhou, De-bao
Zha, Li
Hong, Yang
Jin, Ya-Mei
Lin, Jiao-Jiao
author_facet Zhang, Xin
He, Chuan-Chuan
Liu, Jin-Ming
Li, Hao
Lu, Ke
Fu, Zhi-Qiang
Zhu, Chuan-Gang
Liu, Yi-Ping
Tong, Lai-Bao
Zhou, De-bao
Zha, Li
Hong, Yang
Jin, Ya-Mei
Lin, Jiao-Jiao
author_sort Zhang, Xin
collection PubMed
description BACKGROUND: Schistosomiasis japonica is a common zoonosis. Domestic animals are the primary source of infection and play an important role in disease transmission. The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and, for this reason, diagnostics with a higher sensitivity have become increasingly necessary. It was reported that polymerase chain reaction (PCR)-based methods could be used to detect schistosome infection in humans and animals and presented a high sensitivity and specificity. The present study aimed to develop a PCR-based method for detection of Schistosoma japonicum infection in domestic animals. METHODS: A specific nested-PCR assay was developed to detect S. japonicum infection in domestic animals via amplification of a 231-bp DNA fragment of retrotransposon SjR2. The developed assay was first used in sera and dry blood filter paper (DBFP) from goats and buffaloes at different time points of infection. Then, 78 DBFPs from 39 artificially-infected bovines at 14 and 28 days post-infection and 42 DBFPs from schistosome-negative bovines from the city of Huangshan in the Anhui province were used to evaluate the diagnostic validity. Furthermore, this assay was used to detect S. japonicum infection in domestic animals in Dongzhi and Wangjiang counties. RESULTS: The expected PCR product was detected in eggs and adult worms of S. japonicum and blood samples from S. japonicum-infected goats and water buffaloes, but not from Fasciola and Haemonchus contortus worms. The nested-PCR assay could detect the target S. japonicum DNA in DBFPs from goats and buffaloes after day 3 post-infection. The sensitivity in buffaloes at 14 and 28 days post-infection was 92.30% (36/39) and 100% (39/39), respectively. The specificity was 97.60% (41/42). The positivity rates in Dongzhi and Wangjiang counties were 6.00% and 8.00% in bovines and 22.00% and 16.67% in goats, respectively. The positivity rates in goats in both counties were higher than those in bovines with a significant difference in Dongzhi County but not in Wangjiang County (P < 0.05 and P = 0.23, respectively). CONCLUSIONS: Our results suggest that the developed nested-PCR assay may be used for the diagnosis of S. japonicum infection in domestic animals, and the control of S. japonicum infection in goats should be paid more attention. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40249-017-0298-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-53903782017-04-14 Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals Zhang, Xin He, Chuan-Chuan Liu, Jin-Ming Li, Hao Lu, Ke Fu, Zhi-Qiang Zhu, Chuan-Gang Liu, Yi-Ping Tong, Lai-Bao Zhou, De-bao Zha, Li Hong, Yang Jin, Ya-Mei Lin, Jiao-Jiao Infect Dis Poverty Research Article BACKGROUND: Schistosomiasis japonica is a common zoonosis. Domestic animals are the primary source of infection and play an important role in disease transmission. The prevalence and infectivity of this disease in domestic animals in China have significantly decreased and, for this reason, diagnostics with a higher sensitivity have become increasingly necessary. It was reported that polymerase chain reaction (PCR)-based methods could be used to detect schistosome infection in humans and animals and presented a high sensitivity and specificity. The present study aimed to develop a PCR-based method for detection of Schistosoma japonicum infection in domestic animals. METHODS: A specific nested-PCR assay was developed to detect S. japonicum infection in domestic animals via amplification of a 231-bp DNA fragment of retrotransposon SjR2. The developed assay was first used in sera and dry blood filter paper (DBFP) from goats and buffaloes at different time points of infection. Then, 78 DBFPs from 39 artificially-infected bovines at 14 and 28 days post-infection and 42 DBFPs from schistosome-negative bovines from the city of Huangshan in the Anhui province were used to evaluate the diagnostic validity. Furthermore, this assay was used to detect S. japonicum infection in domestic animals in Dongzhi and Wangjiang counties. RESULTS: The expected PCR product was detected in eggs and adult worms of S. japonicum and blood samples from S. japonicum-infected goats and water buffaloes, but not from Fasciola and Haemonchus contortus worms. The nested-PCR assay could detect the target S. japonicum DNA in DBFPs from goats and buffaloes after day 3 post-infection. The sensitivity in buffaloes at 14 and 28 days post-infection was 92.30% (36/39) and 100% (39/39), respectively. The specificity was 97.60% (41/42). The positivity rates in Dongzhi and Wangjiang counties were 6.00% and 8.00% in bovines and 22.00% and 16.67% in goats, respectively. The positivity rates in goats in both counties were higher than those in bovines with a significant difference in Dongzhi County but not in Wangjiang County (P < 0.05 and P = 0.23, respectively). CONCLUSIONS: Our results suggest that the developed nested-PCR assay may be used for the diagnosis of S. japonicum infection in domestic animals, and the control of S. japonicum infection in goats should be paid more attention. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s40249-017-0298-y) contains supplementary material, which is available to authorized users. BioMed Central 2017-04-13 /pmc/articles/PMC5390378/ /pubmed/28407808 http://dx.doi.org/10.1186/s40249-017-0298-y Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Zhang, Xin
He, Chuan-Chuan
Liu, Jin-Ming
Li, Hao
Lu, Ke
Fu, Zhi-Qiang
Zhu, Chuan-Gang
Liu, Yi-Ping
Tong, Lai-Bao
Zhou, De-bao
Zha, Li
Hong, Yang
Jin, Ya-Mei
Lin, Jiao-Jiao
Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
title Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
title_full Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
title_fullStr Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
title_full_unstemmed Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
title_short Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
title_sort nested-pcr assay for detection of schistosoma japonicum infection in domestic animals
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5390378/
https://www.ncbi.nlm.nih.gov/pubmed/28407808
http://dx.doi.org/10.1186/s40249-017-0298-y
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