Development of In situ hybridization and real-time PCR assays for the detection of Hepatospora eriocheir, a microsporidian pathogen in the Chinese mitten crab Eriocheir sinensis

© 2016 John Wiley & Sons Ltd.

Détails bibliographiques
Publié dans:Journal of fish diseases. - 1998. - 40(2017), 7 vom: 01. Juli, Seite 919-927
Auteur principal: Ding, Z F (Auteur)
Autres auteurs: Chen, J Q, Lin, J, Zhu, X S, Xu, G H, Wang, R L, Meng, Q G, Wang, W
Format: Article en ligne
Langue:English
Publié: 2017
Accès à la collection:Journal of fish diseases
Sujets:Journal Article Eriocheir sinensis Hepatospora eriocheir in situ hybridization real-time PCR
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520 |a A microsporidian parasite, Hepatospora eriocheir, is an emerging pathogen for the Chinese mitten crab Eriocheir sinensis. Currently, there is scant information about the way it transmits infection in the crustacean of commercial importance, including its pathogenesis, propagation and infection route in vivo. In this study, chromogenic in situ hybridization (ISH) and quantitative real-time PCR (qPCR) assays were developed to address this pressing need, and we provided an advance in the detection methods available. Pathogens can be seen in situ with associated lesions using ISH. Positive hybridization signals were noted inside the epithelial cells of the hepatopancreas, and putative free parasite spores were observed within the tubule lumen, which were associated with lesions detected by electron microscopy and haematoxylin and eosin (H&E) analysis. qPCR allows the determination of parasite loads in infected tissues, which is important for understanding disease progression and transmission. The hepatopancreas displayed the biggest statistical copy numbers among different tissues of infected crabs, confirming a tissue-specific pathogen infection characteristic. The qPCR assay also proved to be suitable for the diagnosis of asymptomatic carrier crabs. Combination of the two methods could facilitate the study of H. eriocheir infection mechanism in E. sinensis, enhance the early diagnosis of the pathogen and improve the management of microsporidian diseases in commercial crustaceans 
650 4 |a Journal Article 
650 4 |a Eriocheir sinensis 
650 4 |a Hepatospora eriocheir 
650 4 |a in situ hybridization 
650 4 |a real-time PCR 
700 1 |a Chen, J Q  |e verfasserin  |4 aut 
700 1 |a Lin, J  |e verfasserin  |4 aut 
700 1 |a Zhu, X S  |e verfasserin  |4 aut 
700 1 |a Xu, G H  |e verfasserin  |4 aut 
700 1 |a Wang, R L  |e verfasserin  |4 aut 
700 1 |a Meng, Q G  |e verfasserin  |4 aut 
700 1 |a Wang, W  |e verfasserin  |4 aut 
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