Abstract
We describe a turn-off fluorescence-based strategy for the detection of ATP by making use of aptamer-triggered dsDNA concatamers. This sensitive and easily controlled method is based on consecutive hybridization induced by ATP aptamers and their sectional complementary DNAs to form dsDNA concatamers. The intercalator SYBR Green I (SGI) was employed as a fluorescent probe. In the absence of ATP, the probe produces a strong signal. However, on addition of ATP, the binding of aptamer and ATP cause the concatamers to collapse and to release SGI whose fluorescence then is quenched. The effect was exploited to design a selective ATP assay by relating the decrease in fluorescence to the ATP concentration. A lower detection limit of 6.1 μM and a linear response in the 0 to 5000 μM concentration range was accomplished. The strategy was applied to cellular ATP assays, and the results obtained by this strategy and by the gold standard method are in good agreement. The method is sensitive, simple and cost efficient, and hence is promising in terms of future applications to determine ATP in cellular and other systems.
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Dirks RM, Pierce NA (2004) Triggered amplification by hybridization chain reaction. Proc Natl Acad Sci U S A 101:15275–15278
Chen Y, Xu J, Su J, Xiang Y, Yuan R, Chai YQ (2012) In situ hybridization chain reaction amplification for universal and highly sensitive electrochemiluminescent detection of DNA. Anal Chem 84:7750–7755
Zhang B, Liu BQ, Tang DP, Niessner R, Chen GN, Knopp D (2012) DNA-based hybridization chain reaction for amplified bioelectronic signal and ultrasensitive detection of proteins. Anal Chem 84:5392–5399
Shimron S, Wang F, Orbach R, Willner I (2011) Amplified detection of DNA through the enzyme-free autonomous assembly of hemin/G-quadruplex DNAzyme nanowires. Anal Chem 84:1042–1048
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
Wu JB, Kodzius R, Cao WB, Wen WJ (2014) Extraction, amplification and detection of DNA in microfluidic chip-based assays. Microchim Acta 181:1611–1631
Su C, Liu YF, Ye T, Xiang X, Ji XH, He ZK (2015) Rolling cycle amplification based single-color quantum dots–ruthenium complex assembling dyads for homogeneous and highly selective detection of DNA. Anal Chim Acta 853:495–500
Xia F, White RJ, Zuo X, Patterson A, Xiao Y, Kang D, Gong X, Plaxco KW, Heeger AJ (2010) An electrochemical supersandwich assay for sensitive and selective DNA detection in complex matrices. J Am Chem Soc 132:14346–14348
Shen JW, Li YB, Gu HS, Xia F, Zuo XL (2014) Recent development of sandwich assay based on the nanobiotechnologies for proteins, nucleic acids, small molecules, and ions. Chem Rev 114:7631–7677
Hong CY, Chen X, Liu T, Li J, Yang HH, Chen JH, Chen GN (2013) Ultrasensitive electrochemical detection of cancer-associated circulating microRNA in serum samples based on DNA concatamers. Biosens Bioelectron 50:132–136
Huang JH, Su XF, Li ZG (2012) Enzyme-free and amplified fluorescence DNA detection using bimolecular beacons. Anal Chem 84:5939–5943
Zhao JJ, Chu ZD, Jin X, Zhao SL (2015) A fluorescence polarization assay for nucleic acid based on the amplification of hybridization chain reaction and nanoparticles. Sensors Actuators B Chem 209:116–121
Huang J, Wu YR, Chen Y, Zhu Z, Yang XH, Yang CYJ, Wang KM, Tan WH (2011) Pyrene-excimer probes based on the hybridization chain reaction for the detection of nucleic acids in complex biological fluids. Angew Chem Int Ed 50:401–404
Stubbe J (1990) Ribonucleotide reductases: amazing and confusing. J Biol Chem 265:5329–5332
Fields RD, Burnstock G (2006) Purinergic signalling in neuron–glia interactions. Nat Rev Neurosci 7:423–436
Lin CS, Chen YY, Cai ZX, Zhu Z, Jiang YQ, Yang CYJ, Chen X (2015) A label-free fluorescence strategy for sensitive detection of ATP based on the ligation-triggered super-sandwich. Biosens Bioelectron 63:562–565
Kennedy HJ, Pouli AE, Ainscow EK, Jouaville LS, Rizzuto R, Rutter GA (1999) Glucose generates Sub-plasma membrane ATP microdomains in single islet β-cells: potential role for strategically located mitochondria. J Biol Chem 274:13281–13291
Zhou L, Xue XF, Zhou JH, Li Y, Zhao J, Wu LM (2012) Fast determination of adenosine 5′-triphosphate (ATP) and its catabolites in royal jelly using ultraperformance liquid chromatography. J Agric Food Chem 60:8994–8999
He YF, Liao LF, Xu CH, Wu RR, Li SJ, Yang YY (2015) Determination of ATP by resonance light scattering using a binuclear uranyl complex and aptamer modified gold nanoparticles as optical probes. Microchim Acta 182:419–426
Zyryanov GV, Palacios MA, Anzenbacher PJ (2007) Rational design of a fluorescence-turn-on sensor array for phosphates in blood serum. Angew Chem Int Ed 46:7849–7852
Famulok M, Hartig JS, Mayer G (2007) Functional aptamers and aptazymes in biotechnology, diagnostics, and therapy. Chem Rev 107:3715–3743
Tan WH, Donovan MJ, Jiang JH (2013) Aptamers from cell-based selection for bioanalytical applications. Chem Rev 113:2842–2862
Jiang YN, Liu NN, Guo W, Xia F, Jiang L (2012) Highly-efficient gating of solid-state nanochannels by DNA supersandwich structure containing ATP aptamers: a nanofluidic IMPLICATION logic device. J Am Chem Soc 134:15395–15401
Gao PY, Xia YF, Yang LL, Ma TF, Yang L, Guo QQ, Huang SS (2014) Aptasensor for adenosine triphosphate based on electrode-supported lipid bilayer membrane. Microchim Acta 181:205–212
Huizenga DE, Szostak JW (1995) A DNA aptamer that binds adenosine and ATP. Biochemistry 34:656–665
He XX, Li ZX, Jia XK, Wang KM, Yin JJ (2013) A highly selective sandwich-type FRET assay for ATP detection based on silica coated photon upconverting nanoparticles and split aptamer. Talanta 111:105–110
Kashefi-Kheyrabadi L, Mehrgardi MA (2012) Aptamer-conjugated silver nanoparticles for electrochemical detection of adenosine triphosphate. Biosens Bioelectron 37:94–98
Qiu HZ, Wu NM, Zheng YJ, Chen M, Weng SH, Chen YZ, Lin XH (2015) A robust and versatile signal-on fluorescence sensing strategy based on SYBR green I dye and graphene oxide. Int J Nanomedicine 10:147–156
Acknowledgments
We are thankful for financial support from the National High Technology and Development of China (2012AA022604), the National Natural Science Foundation of China (21405016, 21275028), the Natural Science Foundation of Fujian Province of China (2015 J01043), the Major Program of Medical and Health Foundation of Nanjing Military Region (12Z39), and Program to Young talents of Fujian Province Health System (2013-ZQN-JC-16).
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Qiu, H., Liu, Z., Huang, Z. et al. Aptamer based turn-off fluorescent ATP assay using DNA concatamers. Microchim Acta 182, 2387–2393 (2015). https://doi.org/10.1007/s00604-015-1578-5
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DOI: https://doi.org/10.1007/s00604-015-1578-5