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

doi:10.1007/s00216-019-01651-2

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	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
刊名	ANALYTICAL AND BIOANALYTICAL CHEMISTRY
	2019-04-01
卷号	411 期号:10 页码:2161-2168
关键词	Bacteria detection Flow cytometry Magnetic beads 16S rRNA
ISSN号	1618-2642
DOI	10.1007/s00216-019-01651-2
通讯作者	Zhang, Dun(zhangdun@qdio.ac.cn) ; Qi, Peng(qipeng@qdio.ac.cn)
英文摘要	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.
资助项目	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
语种	英语
出版者	SPRINGER HEIDELBERG
WOS记录号	WOS:000464714400020
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/161143]
专题	海洋研究所_海洋腐蚀与防护研究发展中心
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
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.