Parallel detection of harmful algae using reverse transcription polymerase chain reaction labeling coupled with membrane-based DNA array

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	Parallel detection of harmful algae using reverse transcription polymerase chain reaction labeling coupled with membrane-based DNA array
	Zhang, Chunyun 1,2; Chen, Guofu 1,2,4; Ma, Chaoshuai 1; Wang, Yuanyuan 1; Zhang, Baoyu 1; Wang, Guangce 3
刊名	ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
	2014-03-01
卷号	21 期号:6 页码:4565-4575
关键词	Harmful algal blooms Parallel detection LSU rDNA RT-PCR Membrane-based DNA array
通讯作者	Chen, GF (reprint author), Harbin Inst Technol Weihai, Sch Marine Sci & Technol, Wenhua West Rd 2, Weihai 264209, Shandong, Peoples R China.
英文摘要	Harmful algal blooms (HABs) are a global problem, which can cause economic loss to aquaculture industry's and pose a potential threat to human health. More attention must be made on the development of effective detection methods for the causative microalgae. The traditional microscopic examination has many disadvantages, such as low efficiency, inaccuracy, and requires specialized skill in identification and especially is incompetent for parallel analysis of several morphologically similar microalgae to species level at one time. This study aimed at exploring the feasibility of using membrane-based DNA array for parallel detection of several microalgae by selecting five microaglae, including Heterosigma akashiwo, Chaetoceros debilis, Skeletonema costatum, Prorocentrum donghaiense, and Nitzschia closterium as test species. Five species-specific (taxonomic) probes were designed from variable regions of the large subunit ribosomal DNA (LSU rDNA) by visualizing the alignment of LSU rDNA of related species. The specificity of the probes was confirmed by dot blot hybridization. The membrane-based DNA array was prepared by spotting the tailed taxonomic probes onto positively charged nylon membrane. Digoxigenin (Dig) labeling of target molecules was performed by multiple PCR/RT-PCR using RNA/DNA mixture of five microalgae as template. The Dig-labeled amplification products were hybridized with the membrane-based DNA array to produce visible hybridization signal indicating the presence of target algae. Detection sensitivity comparison showed that RT-PCR labeling (RPL) coupled with hybridization was tenfold more sensitive than DNA-PCR-labeling-coupled with hybridization. Finally, the effectiveness of RPL coupled with membrane-based DNA array was validated by testing with simulated and natural water samples, respectively. All of these results indicated that RPL coupled with membrane-based DNA array is specific, simple, and sensitive for parallel detection of microalgae which shows promise for monitoring natural samples in the future.
学科主题	Environmental Sciences & Ecology
收录类别	SCI
语种	英语
WOS记录号	WOS:000332795700050
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/24141]
专题	海洋研究所_实验海洋生物学重点实验室
作者单位	1.Harbin Inst Technol, Sch Marine Sci & Technol, Weihai 264209, Peoples R China 2.State Ocean Adm, Inst Oceanog 1, Qingdao 266061, Peoples R China 3.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 4.Harbin Inst Technol Weihai, Sch Marine Sci & Technol, Weihai 264209, Shandong, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Chunyun,Chen, Guofu,Ma, Chaoshuai,et al. Parallel detection of harmful algae using reverse transcription polymerase chain reaction labeling coupled with membrane-based DNA array[J]. ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH,2014,21(6):4565-4575.
APA	Zhang, Chunyun,Chen, Guofu,Ma, Chaoshuai,Wang, Yuanyuan,Zhang, Baoyu,&Wang, Guangce.(2014).Parallel detection of harmful algae using reverse transcription polymerase chain reaction labeling coupled with membrane-based DNA array.ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH,21(6),4565-4575.
MLA	Zhang, Chunyun,et al."Parallel detection of harmful algae using reverse transcription polymerase chain reaction labeling coupled with membrane-based DNA array".ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH 21.6(2014):4565-4575.