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

Abstract 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, d...

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Main Authors: Xin Zhang, Chuan-Chuan He, Jin-Ming Liu, Hao Li, Ke Lu, Zhi-Qiang Fu, Chuan-Gang Zhu, Yi-Ping Liu, Lai-Bao Tong, De-bao Zhou, Li Zha, Yang Hong, Ya-Mei Jin, Jiao-Jiao Lin
Format: Article
Language:English
Published: BMC 2017-04-01
Series:Infectious Diseases of Poverty
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Online Access:http://link.springer.com/article/10.1186/s40249-017-0298-y
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author Xin Zhang
Chuan-Chuan He
Jin-Ming Liu
Hao Li
Ke Lu
Zhi-Qiang Fu
Chuan-Gang Zhu
Yi-Ping Liu
Lai-Bao Tong
De-bao Zhou
Li Zha
Yang Hong
Ya-Mei Jin
Jiao-Jiao Lin
author_facet Xin Zhang
Chuan-Chuan He
Jin-Ming Liu
Hao Li
Ke Lu
Zhi-Qiang Fu
Chuan-Gang Zhu
Yi-Ping Liu
Lai-Bao Tong
De-bao Zhou
Li Zha
Yang Hong
Ya-Mei Jin
Jiao-Jiao Lin
author_sort Xin Zhang
collection DOAJ
description Abstract 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.
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spelling doaj.art-288b7b94d87142a791c4e7ae97e863de2022-12-21T23:58:21ZengBMCInfectious Diseases of Poverty2049-99572017-04-01611710.1186/s40249-017-0298-yNested-PCR assay for detection of Schistosoma japonicum infection in domestic animalsXin Zhang0Chuan-Chuan He1Jin-Ming Liu2Hao Li3Ke Lu4Zhi-Qiang Fu5Chuan-Gang Zhu6Yi-Ping Liu7Lai-Bao Tong8De-bao Zhou9Li Zha10Yang Hong11Ya-Mei Jin12Jiao-Jiao Lin13Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureAnhui Center for Animal Disease Control and PreventionWangjiang county Center for Animal Husbandry and Veterinary BureauDongzhi county Center for Animal Husbandry and Veterinary BureauHuangshan Center for Animal Disease Control and PreventionShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureShanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology, Ministry of AgricultureAbstract 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.http://link.springer.com/article/10.1186/s40249-017-0298-ySchistosomiasisDomestic animalsNested-PCRDiagnosis
spellingShingle Xin Zhang
Chuan-Chuan He
Jin-Ming Liu
Hao Li
Ke Lu
Zhi-Qiang Fu
Chuan-Gang Zhu
Yi-Ping Liu
Lai-Bao Tong
De-bao Zhou
Li Zha
Yang Hong
Ya-Mei Jin
Jiao-Jiao Lin
Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animals
Infectious Diseases of Poverty
Schistosomiasis
Domestic animals
Nested-PCR
Diagnosis
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 Schistosomiasis
Domestic animals
Nested-PCR
Diagnosis
url http://link.springer.com/article/10.1186/s40249-017-0298-y
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