Combination of a flow cytometric bead system with 16S rRNA-targeted oligonucleotide probes for bacteria detection

doi:10.1007/s00216-019-01651-2

	Combination of a flow cytometric bead system with 16S rRNA-targeted oligonucleotide probes for bacteria detection
	Zeng, Yan1,2,3 ; Zhang, Dun 1,2,3; Qi, Peng1,2,3
	2019-04-01
发表期刊	ANALYTICAL AND BIOANALYTICAL CHEMISTRY
ISSN	1618-2642
卷号	411 期号:10 页码:2161-2168
通讯作者	Zhang, Dun([email protected]) ; Qi, Peng([email protected])
摘要	Here we report a bacteria detection method based on a flow cytometric bead system and 16S rRNA-targeted oligonucleotide probes. Polymerase chain reaction (PCR) was first used to acquire bacterial DNA including bacteria-specific sequences. Half of the resulting target DNA was then captured by a capture probe immobilized on a magnetic microbead (MB) surface. The other half of the target DNA was hybridized with a fluorescence-labeled signal probe. In this manner, a sandwich DNA hybridization involving a MB-based capture probe, the target DNA, and a signal probe was realized. The MB carriers modified with reporter dye were analyzed one by one by flow cytometry through a capillary. Using PCR amplicons and this flow cytometric bead system, a detection limit of 180 cfu mL(-1) was achieved, along with high selectivity that permitted the discrimination of different targets when challenged with control bacteria targets and multiplexing capabilities that enabled the simultaneous detection of two kinds of bacteria. Given these advantages, the developed method can be used for the highly sensitive and specific PCR amplicon analysis of DNA extracted from a fresh bacterial culture, as well as multiplex target analysis.
关键词	Bacteria detection Flow cytometry Magnetic beads 16S rRNA
DOI	10.1007/s00216-019-01651-2
收录类别	SCI
语种	英语
资助项目	National Natural Science Foundation of China[41876101]###507; China Postdoctoral Science Foundation[2016M602199]###2528; Young Elite Scientists Sponsorship Program by CAST[2018QNRC001]###508; Applied Basic Research Program of Nantong[GY12017005]###509; AoShan Talent Program###510; Qingdao National Laboratory for Marine Science and Technology###117; National Natural Science Foundation of China[41876101]; China Postdoctoral Science Foundation[2016M602199]; Young Elite Scientists Sponsorship Program by CAST[2018QNRC001]; Applied Basic Research Program of Nantong[GY12017005]; AoShan Talent Program; Qingdao National Laboratory for Marine Science and Technology
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemical Research Methods ; Chemistry, Analytical
WOS记录号	WOS:000464714400020
出版者	SPRINGER HEIDELBERG
引用统计	被引频次：8[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/161144
专题	海洋环境腐蚀与生物污损重点实验室
通讯作者	Zhang, Dun; Qi, Peng
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Marine Environm Corros & Biofouling, 7 Nanhai Rd, Qingdao 266071, Shandong, Peoples R China 2.Chinese Acad Sci, Ctr Ocean Megasci, 7 Nanhai Rd, Qingdao 266071, Shandong, Peoples R China 3.Qingdao Natl Lab Marine Sci & Technol, Open Studio Marine Corros & Protect, 1 Wenhai Rd, Qingdao 266237, Shandong, Peoples R China
第一作者单位	中国科学院海洋研究所; 中国科学院海洋大科学研究中心
通讯作者单位	中国科学院海洋研究所; 中国科学院海洋大科学研究中心
推荐引用方式 GB/T 7714	Zeng, Yan,Zhang, Dun,Qi, Peng. Combination of a flow cytometric bead system with 16S rRNA-targeted oligonucleotide probes for bacteria detection[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2019,411(10):2161-2168.
APA	Zeng, Yan,Zhang, Dun,&Qi, Peng.(2019).Combination of a flow cytometric bead system with 16S rRNA-targeted oligonucleotide probes for bacteria detection.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,411(10),2161-2168.
MLA	Zeng, Yan,et al."Combination of a flow cytometric bead system with 16S rRNA-targeted oligonucleotide probes for bacteria detection".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 411.10(2019):2161-2168.

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