Nested-PCR assay for detection of Schistosoma japonicum infection in domestic animalsReportar como inadecuado




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Infectious Diseases of Poverty

, 6:86

Schistosomiasis Research: Providing the Tools Needed for Elimination

Abstract

BackgroundSchistosomiasis 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.

MethodsA 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.

ResultsThe 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.

ConclusionsOur 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.

KeywordsSchistosomiasis Domestic animals Nested-PCR Diagnosis AbbreviationsDBFPDry blood filter paper

IHAIndirect hemagglutination assay

MHTMiracidium hatching test

PBSPhosphate buffered saline

PCRPolymerase chain reaction

SEASoluble egg antigen

SWASoluble worm antigens

Electronic supplementary materialThe 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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Autor: Xin Zhang - Chuan-Chuan He - Jin-Ming Liu - Hao Li - Ke Lu - Zhi-Qiang Fu - Chuan-Gang Zhu - Yi-Ping Liu - Lai-Bao Tong -

Fuente: https://link.springer.com/







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