IOCAS-IR  > 海洋环境腐蚀与与生物污损重点实验室
Combination of a flow cytometric bead system with 16S rRNA-targeted oligonucleotide probes for bacteria detection
Zeng, Yan1,2,3; Zhang, Dun1,2,3; Qi, Peng1,2,3
2019-04-01
Source PublicationANALYTICAL AND BIOANALYTICAL CHEMISTRY
ISSN1618-2642
Volume411Issue:10Pages:2161-2168
Corresponding AuthorZhang, Dun(zhangdun@qdio.ac.cn) ; Qi, Peng(qipeng@qdio.ac.cn)
AbstractHere 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.
KeywordBacteria detection Flow cytometry Magnetic beads 16S rRNA
DOI10.1007/s00216-019-01651-2
Indexed BySCI
Language英语
Funding ProjectNational 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 Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS IDWOS:000464714400020
PublisherSPRINGER HEIDELBERG
Citation statistics
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/161144
Collection海洋环境腐蚀与与生物污损重点实验室
Corresponding AuthorZhang, Dun; Qi, Peng
Affiliation1.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
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Corresponding Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
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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