The use of beta-tubulin gene for phylogenetic analysis of the microsporidian parasite Enterocytozoon hepatopenaei (EHP) and in the development of a nested PCR as its diagnostic tool

文献类型： 外文期刊

第一作者： Han, Jee Eun

作者： Han, Jee Eun;Tang, Kathy F. J.;Kim, Ji Hyung

作者机构：

关键词： Shrimp disease; Enterocytozoon hepatopenaei; beta-Tubulin; Nested PCR

期刊名称：AQUACULTURE （ 影响因子：4.242； 五年影响因子：4.723 ）

ISSN： 0044-8486

年卷期： 2018 年 495 卷

页码：

收录情况： SCI

摘要： Tubulins are major components of microtubules, structures that are involved in many cellular processes. The gene beta-tubulin, which encodes a beta-tubulin protein, is conserved, and yet with sufficient variability to be useful for phylogenetic studies and for use in molecular diagnostic protocols. We sequenced and analyzed a fragment (870-bp) of beta-tubulin gene of Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp and associated with growth retardation. The sequence was then used to conduct a phylogenetic study to determine the position of EHP within the Microsporidia and to develop primers for an EHP nested PCR method. Blast analysis revealed a 75% nucleotide identity of EHP beta-tubulin to that of E. bieneusi. Similar to other studies based on the small subunit rRNA sequence, the phylogenetic analysis using beta-tubulin sequences clustered EHP with E. bieneusi and two other crustacean microsporidia, Agmasoma penaeii and Hepatospora eriocheir. As for the diagnostic use, a nested PCR method was developed that successfully detected EHP in infected Penaeus vannamei from Indonesia, Thailand, Vietnam, and India and in some cases of Artemia biomass. There were no cross-reactions to the shrimp and other crustacean genomes. This nested PCR is 100-fold more sensitive than the onestep PCR and thus suitable for detecting a low level of EHP infection.

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