其他题名Development a rapid assay to detect the edible jellyfish Rhopilema esculentum larvae using loop-mediated isothermal amplification method
孙婷婷; 刘中援; 梁丽琨; 董志军
发表期刊应用海洋学学报
ISSN2095-4972
2017-11-15
卷号36期号:4页码:538-544
关键词marine biologyRhopilema esculentumloop-mediated isothermal amplification methodrapid detectionpolyps海洋生物学海蜇环介导等温扩增快速检测螅状幼体
DOIDOI:10.3969/J.ISSN.2095-4972.2017.04.010
产权排序(1) 烟台大学生命科学学院; (2) 中国科学院烟台海岸带研究所
作者部门海岸带生物资源高效利用研究与发展中心
英文摘要It is difficult to identify the scyphomedusae larvae by traditional morphological methods,due to the complex morphological characteristics in different stages of their life cycle. For early identifying and detecting of this species,a simple and effective method using the loop-mediated isothermal amplification method (LAMP) has been developed. LAMP primers of Rhopilema esculentum were designed based on the mitochondrial cytochrome c oxidase subunit Ⅰ (COⅠ) gene. The amplification conditions,including the betaine and dNTPs concentrations,the inner primer to outer primer concentration ratio,reaction time and temperature,were optimized. The specificity,sensitivity and repeatability of the method were also tested. Our results show that the primers had specificity and repeatability. Samples from different geographic populations of R. esculentum and polyps produced positive results,while the other non-target scyphomedusae species produced negative results. In addition,the LAMP assay was more sensitive than conventional PCR. The detection limit of LAMP method was 0. 1 ng DNA. Therefore,the established LAMP assay is a sensitive,specific,fast,and easily performed method for detection of the edible jellyfish R. esculentum.
中文摘要针对海蜇早期生活史阶段幼体种类鉴定困难的问题,建立一种快速、灵敏、操作简便的环介导等温扩增(LAMP)快速检测方法.根据海蜇的线粒体细胞色素C氧化酶Ⅰ基因(COⅠ)编码序列,设计 LAMP引物,建立了海蜇幼体LAMP快速检测方法,并对LAMP反应体系、反应温度和反应时间进行了优化.结果表明,LAMP方法对海蜇目的基因的最小检测限为0. 1 ng DNA.该检测方法特异性较强,与黄渤海其他常见钵水母种类海月水母、白色霞水母、沙海蜇均无交叉反应.利用本研究建立的海蜇 LAMP检测方法对不同海域采集的成体海蜇样品和海蜇螅状幼体进行检测,结果显示,成体海蜇和海蜇幼体均呈现阳性结果,表明海蜇LAMP检测方法可以对海蜇幼体进行快速有效的检测.海蜇LAMP检测方法简单快速、灵敏且特异性强,可使其运用于沿海渔业部门海蜇资源的调查工作.
文章类型Article
收录类别CSCD
语种中文
关键词[WOS]Oceanography
研究领域[WOS]Oceanography; 海洋学
CSCD记录号CSCD:6118554
引用统计
文献类型期刊论文
条目标识符http://ir.yic.ac.cnhttp://ir.yic.ac.cn/handle/133337/24274
专题海岸带生物学与生物资源利用重点实验室_海岸带生物资源高效利用研究与发展中心
作者单位1.烟台大学生命科学学院;
2.中国科学院烟台海岸带研究所
推荐引用方式
GB/T 7714孙婷婷,刘中援,梁丽琨,等. 海蜇幼体环介导等温扩增快速检测方法的建立和应用[J]. 应用海洋学学报,2017,36(4):538-544.
APA孙婷婷,刘中援,梁丽琨,&董志军.(2017).海蜇幼体环介导等温扩增快速检测方法的建立和应用.应用海洋学学报,36(4),538-544.
MLA孙婷婷,et al."海蜇幼体环介导等温扩增快速检测方法的建立和应用".应用海洋学学报 36.4(2017):538-544.
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