In recent years,bonamiosis has frequently occurred in European areas,which has caused the death of oyster in a wide range and brought enormous economic losses to the breeding industry of oyster. Nowadays,the study on ...In recent years,bonamiosis has frequently occurred in European areas,which has caused the death of oyster in a wide range and brought enormous economic losses to the breeding industry of oyster. Nowadays,the study on Bonamia sp. is still in the elementary phase. The technology of pathogen’s culture in vitro is the basis for further study on the pathogenesis of Bonamia sp.,its interaction with hosts and the prevention and control of the related disease. In this study,total tissues of oyster identified by PCR were used as culture media to in vitro culture. After one month,they were identified by the method of in situ hybridization. It was found that the results of in situ hybridization were accordant with PCR results. And Bonamia ostreae were detected by in situ hybridization after B. ostreae were cultured for one month. We successfully established a simple and feasible method for in vitro culturing B. ostreae.展开更多
应用聚合酶链式反应(PCR)方法对福建、广东和海南等地沿海的养殖牡蛎进行包拉米虫、派琴虫和单孢子虫检测.结果表明,这些地区的牡蛎均不同程度地感染这些原虫,经鉴定病原为牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫.根据基因库中奥尔...应用聚合酶链式反应(PCR)方法对福建、广东和海南等地沿海的养殖牡蛎进行包拉米虫、派琴虫和单孢子虫检测.结果表明,这些地区的牡蛎均不同程度地感染这些原虫,经鉴定病原为牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫.根据基因库中奥尔森派琴虫和尼氏单孢子虫的基因序列设计多对特异性引物,检测包拉米虫的引物采用世界动物卫生组织推荐引物,通过对多重PCR反应条件的优化,建立可同时检测这3种原虫的多重PCR方法.运用该方法对样品中的牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫进行扩增,结果得到与试验设计相符的303、480和749 bp 3条特异性扩增条带,对其他贝类病原核酸的扩增均为阴性.多重PCR方法最低能检测到10 pg牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫DNA,表明该方法适用于这3种原虫的快速检测和鉴别诊断.展开更多
基金Supported by National Science and Technology Supporting Program(2012BAK11B04)
文摘In recent years,bonamiosis has frequently occurred in European areas,which has caused the death of oyster in a wide range and brought enormous economic losses to the breeding industry of oyster. Nowadays,the study on Bonamia sp. is still in the elementary phase. The technology of pathogen’s culture in vitro is the basis for further study on the pathogenesis of Bonamia sp.,its interaction with hosts and the prevention and control of the related disease. In this study,total tissues of oyster identified by PCR were used as culture media to in vitro culture. After one month,they were identified by the method of in situ hybridization. It was found that the results of in situ hybridization were accordant with PCR results. And Bonamia ostreae were detected by in situ hybridization after B. ostreae were cultured for one month. We successfully established a simple and feasible method for in vitro culturing B. ostreae.
文摘应用聚合酶链式反应(PCR)方法对福建、广东和海南等地沿海的养殖牡蛎进行包拉米虫、派琴虫和单孢子虫检测.结果表明,这些地区的牡蛎均不同程度地感染这些原虫,经鉴定病原为牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫.根据基因库中奥尔森派琴虫和尼氏单孢子虫的基因序列设计多对特异性引物,检测包拉米虫的引物采用世界动物卫生组织推荐引物,通过对多重PCR反应条件的优化,建立可同时检测这3种原虫的多重PCR方法.运用该方法对样品中的牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫进行扩增,结果得到与试验设计相符的303、480和749 bp 3条特异性扩增条带,对其他贝类病原核酸的扩增均为阴性.多重PCR方法最低能检测到10 pg牡蛎包拉米虫、奥尔森派琴虫和尼氏单孢子虫DNA,表明该方法适用于这3种原虫的快速检测和鉴别诊断.
