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使用与核酸酶保护分析相结合的夹心杂交法(NPA-SH)检测球形棕囊藻。

Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protection assay (NPA-SH).

作者信息

Zhen Yu, Mi Tiezhu, Yu Zhigang

机构信息

Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China.

出版信息

J Environ Sci (China). 2008;20(12):1481-6. doi: 10.1016/s1001-0742(08)62553-x.

Abstract

Phaeocystis globosa Scherffel is one of the common harmful algae species in coastal waters of the southeastern China. In this study, sandwich hybridization integrated with nuclease protection assay (NPA-SH) was used to qualitatively and quantitatively detect P. globosa. Results showed that this method had good applicability and validity in analyzing the samples from laboratory cultures and from fields. The linear regression equation for P. globosa was obtained, and the lowest detection number of cells was 1.8 x 10(4) cells. Statistics showed that there was no distinct difference between the results of detecting the microalgae by NPA-SH and traditional microscopy. This technique has good reliability, accuracy, and can give a remarkably high sample processing rate. Sandwich hybridization integrated with nuclease protection assay will provide an efficient alternative to microscopic method for monitoring and investigating the bloom of P. globosa.

摘要

球形棕囊藻(Phaeocystis globosa Scherffel)是中国东南沿海常见的有害藻类之一。本研究采用核酸酶保护分析-夹心杂交法(NPA-SH)对球形棕囊藻进行定性和定量检测。结果表明,该方法在分析实验室培养样本和野外样本时具有良好的适用性和有效性。得到了球形棕囊藻的线性回归方程,最低细胞检测数量为1.8×10⁴个细胞。统计结果表明,NPA-SH检测微藻的结果与传统显微镜检测结果无显著差异。该技术具有良好的可靠性、准确性,且样本处理速度极快。核酸酶保护分析-夹心杂交法将为监测和调查球形棕囊藻水华提供一种有效的替代显微镜检测的方法。

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