广东海洋大学学报2016，Vol.36Issue(4)：50-54,5.DOI:10.3969/j.issn.1673-9159.2016.04.009
凡纳滨对虾肠道上皮细胞微胞子虫PCR检测方法的建立与应用
Development and Application ofa PCR DetectionAssayof Microsporidium Enterocytozoon hepatopenaei in Pacific WhiteLeg Shrimp Litopenaeus vannamei
摘要
Abstract
A pair of specific primers was designed, and a rapid PCR method was established for detection ofmicrosporidiumEnterocytozoonhepatopenaeiin Pacific white leg shrimpLitopenaeus vannameiaccording to the publishedgenesequences in GenBank by optimization of the reaction parameters.The specific band of 330bp were amplified from the positive shrimps, but no specific band wasfound fromshrimps with white spotsyndrome virus (WSSV), Taura syndrome virus (TSV), infectious hypodermal and hematopoietic necrosis virus (IHHNV), Baculovirus penaei (BP), and Pleistophora-like microsporidium,Myxosporeaand healthy shrimps.According to theSequencing analysis, thesequence ofPCR product had99.8%homology with that of EHP gene in GenBank which indicated the highaccuracy ofthis method. The sensitivity test of this method showed thatthe lowest detectable limitDNA plasmidof themicrosporidawas100 fg. 755 clinicalshrimpsamples collectedfromprovinces ofGuangdong,Guangxi, Jiangsu, Shandong, andHainanwere detected by this method, and then 21positive samples were found. Thefindings indicated that the PCR assay could be usedinthe detection ofPacific white leg shrimpsamples.关键词
凡纳滨对虾/肠道上皮细胞微胞子虫/PCR检测/临床应用Key words
Litopenaeus vannamei/Enterocytozoon hepatopenaei/PCR detection/clinical application分类
农业科技引用本文复制引用
雷燕,肖洋,张会军,王娟,张文文,卢刚,王雪鹏..凡纳滨对虾肠道上皮细胞微胞子虫PCR检测方法的建立与应用[J].广东海洋大学学报,2016,36(4):50-54,5.基金项目
山东省现代农业产业技术体系（SDAIT-19）；泰安市科技发展计划项目（201440774）；浙江省近岸水域生物资源开发与保护重点实验室开放基金项目 （）
