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A cytometric assay for ultrasensitive and robust detection of human telomerase RNA based on toehold strand displacement. 一种基于脚端链位移的超灵敏和稳健检测人类端粒酶RNA的细胞分析方法。

IF 12.6

Biosensors & bioelectronics Pub Date : 2017-01-15 Epub Date: 2016-08-13 DOI: 10.1016/j.bios.2016.08.038

Jing Xu, Yanjun Wang, Luzhu Yang, Yanfang Gao, Baoxin Li, Yan Jin

{"title":"A cytometric assay for ultrasensitive and robust detection of human telomerase RNA based on toehold strand displacement.","authors":"Jing Xu, Yanjun Wang, Luzhu Yang, Yanfang Gao, Baoxin Li, Yan Jin","doi":"10.1016/j.bios.2016.08.038","DOIUrl":"https://doi.org/10.1016/j.bios.2016.08.038","url":null,"abstract":"Human telomerase RNA (hTR), works as a template for synthesis of telomeric DNA repeats at the ends of linear eukaryotic chromosomes, is overexpressed in tumor cells and its concentration has a positive correlation with telomerase activity. The lack of facile and reliable method for detection of hTR in complex matric limited its application for clinical diagnosis. To address the limitation, herein, we proposed a facile and reliable flow cytometric assay for sensitive and specific detection of hTR by combing magnetic enrichment with signal amplification of DNA toehold strand displacement reaction (TSDR). Two hairpin DNA probes of TSDR are ingeniously designed, including biotinylated hairpin DNA1 (H1) and carboxyfluorescein (FAM)-labeled hairpin DNA2 (F-H2). Firstly, H1 was immobilized on streptavidin-functionalized magnetic beads (STV-MBs) through biotin-avidin interaction. In the presence of hTR DNA, TSDR between H1 and F-H2 was triggered to continuously form H1/H2 duplex, resulting in a \"turn on\" fluorescence on the surface of MBs. Due to fluorescence amplification of TSDR and magnetic enrichment, hTR-DNA can be sensitively, specifically and facile analyzed by flow cytometry and fluorescence microscopy imaging. The detection limit of flow cytometry is 0.3pM, which is superior to those of most existing approaches. Moreover, the proposed strategy can be successfully utilized to detect hTR in complex biological media as well. Therefore, an enzyme-free amplification approach is provided for robust and rapid detecting hTR DNA, which offers a facile, reliable and sensitive method for studying disease-related gene.","PeriodicalId":502209,"journal":{"name":"Biosensors & bioelectronics","volume":" ","pages":"1071-1076"},"PeriodicalIF":12.6,"publicationDate":"2017-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bios.2016.08.038","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39975471","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 22

Bacteria adsorption on hydrophilic surfaces for the sensitive detection of pathogenic bacteria using a single tube chamber system. 细菌在亲水性表面的吸附用于病原菌的单管室系统的敏感检测。

IF 12.6

Biosensors & bioelectronics Pub Date : 2010-12-15 Epub Date: 2010-08-20 DOI: 10.1016/j.bios.2010.08.037

Ji Yeong Won, Junhong Min, Jung-Hwan Park

{"title":"Bacteria adsorption on hydrophilic surfaces for the sensitive detection of pathogenic bacteria using a single tube chamber system.","authors":"Ji Yeong Won, Junhong Min, Jung-Hwan Park","doi":"10.1016/j.bios.2010.08.037","DOIUrl":"https://doi.org/10.1016/j.bios.2010.08.037","url":null,"abstract":"Here, we developed a simple and effective bacterial isolation method that can be directly used for the detection of pathogenic bacteria. This approach only requires a single plastic tube chamber that can performing serial processes such as cell gathering, cell lysis, nucleic acid amplification, and signaling without the need to transfer samples from one chamber to another. A TEOS (tetraethoxysilane) surface was selected for this application because of its superior performance in the amplification process as well as the ability of bacteria to adsorb to its surface, which is necessary for all processes to be performed in single chamber. The optimal aquatic buffer conditions for bacteria adsorption on the hydrophilic surface were determined to be 1% polyethylene glycerol (PEG) and 10 mM MgCl(2) in 100 mM phosphate at pH 4 for the gram negative bacteria, Escherichia coli O157:H7 (E. coli O157:H7) and 10 mM Na(2)SO(4) in 100 mM phosphate in 100 mM phosphate at pH 4 for the gram positive bacteria, Bacillus cereus (B. cereus). When these divalent cation and anion (MgSO(4)) containing acidic solutions were used, 40% of both bacteria adsorbed onto the hydrophilic surface at a loading rate of 2 mL/min after introduction of low concentrations of bacteria. This method was directly employed to detect E. coli O157:H7 in beef using a single plastic tube chamber that was partially filled with nickel micro beads coated with TEOS. In this system, E. coli O157:H7 were lysed by induction heating of the nickel micro beads. The extracted mRNA was readily amplified and detected by adding an isothermal amplification mixture (NASBA, nucleic acid sequence based amplification) containing a hair-loop type reporting probe with FAM and DABCYL. As a result, this highly sensitive sensing tool could detect very low concentrations of E. coli [10(0) CFU/1 g of beef].","PeriodicalId":502209,"journal":{"name":"Biosensors & bioelectronics","volume":" ","pages":"1763-7"},"PeriodicalIF":12.6,"publicationDate":"2010-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bios.2010.08.037","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40075247","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 17

Biological investigation using a shear horizontal surface acoustic wave sensor: small "click generated" DNA hybridization detection. 生物学研究用剪切水平表面声波传感器:小“点击产生”DNA杂交检测。

IF 12.6

Biosensors & bioelectronics Pub Date : 2010-12-15 Epub Date: 2010-08-19 DOI: 10.1016/j.bios.2010.08.036

Chouki Zerrouki, Najla Fourati, Romain Lucas, Julien Vergnaud, Jean-Marie Fougnion, Rachida Zerrouki, Christine Pernelle

{"title":"Biological investigation using a shear horizontal surface acoustic wave sensor: small \"click generated\" DNA hybridization detection.","authors":"Chouki Zerrouki, Najla Fourati, Romain Lucas, Julien Vergnaud, Jean-Marie Fougnion, Rachida Zerrouki, Christine Pernelle","doi":"10.1016/j.bios.2010.08.036","DOIUrl":"https://doi.org/10.1016/j.bios.2010.08.036","url":null,"abstract":"We have used a 104 MHz lithium tantalate (LiTaO(3)) surface acoustic wave (SAW) sensor to investigate DNA probes grafting and their further hybridization with natural and click generated (Cg-DNA) oligonucleotides. Natural DNA targets of different strand lengths, tosyl-di(tri, tetra) thymidine and azido-di(tri, tetra) thymidine oligonucleotides were tested. In our case, and besides the follow-up of a 34mer DNA hybridization, we detected complementarity between natural DNA probes and azido-tetra-thymidine for the first time, whereas previous hybridization studies reported a minimal of 10-mer oligonucleotides recognition length. We also demonstrated that contrarily to natural DNA, the synthesized oligonucleotides present stable bonds with complementary DNA strands. Frequency responses of both grafting and hybridization have shown the same shape: an exponential decay with different time constants, (187±1)s and (68±19) s for grafting and hybridization respectively. We have also shown that recognition between DNA strands and tetranucleotide analogues is comparable to natural 34mer DNA bases and presents the same time constant within uncertainties.","PeriodicalId":502209,"journal":{"name":"Biosensors & bioelectronics","volume":" ","pages":"1759-62"},"PeriodicalIF":12.6,"publicationDate":"2010-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bios.2010.08.036","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40062361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 17