| 实时荧光定量 PCR 法快速检测赤潮环状异帽藻 |
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| 引用本文： | 石彦红,张凤英,马凌波. 实时荧光定量 PCR 法快速检测赤潮环状异帽藻[J]. 中国水产科学, 2010, 17(2): 267-273 |
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| 作者姓名： | 石彦红 张凤英 马凌波 |
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| 作者单位： | 1. 上海海洋大学,生命科学与技术学院,上海,201306;中国水产科学研究院,东海水产研究所,农业部海洋与河口渔业资源及生态重点开放实验室,上海,200090
2. 中国水产科学研究院,东海水产研究所,农业部海洋与河口渔业资源及生态重点开放实验室,上海,200090 |
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| 基金项目： | 中央级公益性科研院所基本科研业务费专项资金(中国水产科学研究院东海水产研究所)资助项目，上海市科学技术委员会科研计划项目 |
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| 摘 要： | 以环状异帽藻(Heterocapsa circularisquama)ITS1区为靶序列设计了一对特异性引物,运用Real-time PCR方法,对环状异帽藻进行了定性定量检测。以其他9种赤潮微藻为对照验证其特异性。结果表明,仅含环状异帽藻DNA模板的样品有扩增,其他藻没有检测到扩增信号。以超声波破碎得到的藻液为标准品,所得标准曲线具有高度线性相关,相关系数R2为0.989,并且可以在30个循环内检测到0.5个细胞。对在不同培养条件下获得的环状异帽藻DNA样品进行检测,都能获得扩增信号。利用该方法建立的标准曲线对初步模拟现场样品进行定量检测,与显微镜计数结果基本一致。
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| 关 键 词： | 环状异帽藻 鉴定 定量检测 |
| 修稿时间： | 2010-03-30 |
Development of a real-time PCR assay for rapid detection and quantification of Heterocapsa circularisquama |
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| SHI Yanhong,ZHANG Fengying,MA Lingbo. Development of a real-time PCR assay for rapid detection and quantification of Heterocapsa circularisquama[J]. Journal of Fishery Sciences of China, 2010, 17(2): 267-273 |
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| Authors: | SHI Yanhong ZHANG Fengying MA Lingbo |
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| Affiliation: | SHI Yanhong 1,2,ZHANG Fengying 2,MA Lingbo 1,2 (1.College of Fisheries , Life Science,Shanghai Ocean University,Shanghai 201306,China,2.East China Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Key Laboratory of Marine , Estuarine Fisheries Resources , Ecology,Ministry of Agriculture,Shanghai 200090,China) |
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| Abstract: | A real-time polymerase chain reaction(PCR)assay was designed and evaluated for rapid detection and quantification of the toxic algae Heterocapsa circularisquama.A pair of specific primers was designed from the sequence of ITS1 region,of which the PCR specificity was examined compared with the other nine dinoflagellates.PCR amplifications were detected only from samples which contained H.circularisquama cells,and specific signals were not detected from any other microalgae.Based on crushed algae by ultrasoun... |
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| Keywords: | Real-time PCR ITS1 Heterocapsa circularisquama real-time PCR ITS1 quantitive identification |
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