Corresponding Author

Xiao-hua ZHANG(mickyxie@hnu.edu.cn);
Jin-hua CHEN(chenjinhua@hnu.edu.cn)


A simple “signal-on” photoelectrochemical (PEC) sensing platform for sensitive assay of nucleic acids was developed by coupling catalytic hairpin assembly (CHA) signal amplification strategy with Ru(NH3)63+. Herein, cadmium sulfide (CdS) was deposited on the TiO2/indium tin oxide (ITO) electrode by a method of successive ionic layer adsorption and reaction (SILAR), serving as one kind of photoelectric material to broaden absorption range of TiO2 and to improve the photoelectric conversion efficiency. Thereafter, the capture DNA (C-DNA) was immobilized on the CdS/TiO2/ITO electrode. Simultaneously, Au-hairpin DNA probe 1 (Au-HP1) and hairpin DNA probe 2 (HP2) were able to hybridize to produce many Au-HP1:HP2 complexes under the existence of target DNA (T-DNA) based on CHA process. After that, C-DNA could capture Au-HP1:HP2 complex, leading to lots of double-stranded DNAs on the electrode to load numerous Ru(NH3)63+, which resulted in a remarkable increase of photocurrent. As a result, a wide linear range (10 fmol·L-1 to 1500 fmol·L-1) and a low detection limit (6.19 fmol·L-1) toward T-DNA were achieved. The developed method would have great potential applications in bioanalysis, screening of new drugs and early disease diagnosis.

Graphical Abstract


photoelectrochemical biosensor, nucleic acid, catalytic hairpin assembly, [Ru(NH3)6]Cl3, CdS, TiO2

Publication Date


Online Available Date


Revised Date


Received Date



[1] Boulikas T. Common structural features of replication origins in all life forms[J]. Journal of Cellular Biochemistry, 1996, 60(3): 297-316.
[2] Orgel L E. The origin of life-a review of facts and speculations[J]. Trends in Biochemical Sciences, 1998, 23(12): 491-495.
[3] Baeissa A, Dave N, Smith B D, et al. DNA-functionalized monolithic hydrogels and gold nanoparticles for colorimetric DNA detection[J]. ACS Applied Materials and Interfaces,
2010, 2(12): 3594-3600.
[4] Zhang Y Y, Tang Z W, Wang J, et al. Hairpin DNA switch for ultrasensitive spectrophotometric detection of DNA hybridization based on gold nanoparticles and enzyme signal
amplification[J]. Analytical Chemistry, 2010, 82(15):6440-6446.
[5] Gao Y, Li B X. G-quadruplex DNAzyme-based chemiluminescence biosensing strategy for ultrasensitive DNA detection: combination of exonuclease III-assisted signal
amplification and carbon nanotubes-assisted background reducing[J]. Analytical Chemistry, 2013, 85(23): 11494-11500.
[6] Gao Y, Li B X. Exonuclease III-assisted cascade signal amplification strategy for label-free and ultrasensitive chemiluminescence detection of DNA[J]. Analytical Chemistry,
2014, 86(17): 8881-8887.
[7] Xiong E H, Yan X X, Zhang X H, et al. Exonuclease IIIassisted cascade signal amplification strategy for labelfree and ultrasensitive electrochemical detection of nucleic
acids[J]. Biosensors and Bioelectronics, 2017, 87: 732-736.
[8] Xiong E H, Zhang X H, Liu Y Q, et al. Ultrasensitive electrochemical detection of nucleic acids based on the dualsignaling electrochemical ratiometric method and exonuclease
III-assisted target recycling amplification strategy[J]. Analytical Chemistry, 2015, 87(14): 7291-7296.
[9] Xiong E, Li Z Z, Zhang X H, et al. Triple-helix molecular switch electrochemical ratiometric biosensor for ultrasensitive detection of nucleic acids[J]. Analytical Chemistry,
2017, 89(17): 8830-8835.
[10] Zhang H R, Xu J J, Chen H Y. Electrochemiluminescence ratiometry: a new approach to DNA biosensing[J].Analytical Chemistry, 2013, 85(11): 5321-5325.
[11] Chai Y, Tian D Y, Wang W, et al. A novel electrochemiluminescence strategy for ultrasensitive DNA assay using luminol functionalized gold nanoparticles multi-labeling and amplification of gold nanoparticles and biotin-streptavidin system[J]. Chemical Communications, 2010, 46(40): 7560-7562.
[12] Huang J H, Su X F, Li Z G. Enzyme-free and amplified fluorescence DNA detection using bimolecular beacons [J]. Analytical Chemistry, 2012, 84(14): 5939-5943.
[13] Liu G, Li J, Feng D Q, et al. Silver nanoclusters beacon as stimuli-responsive versatile platform for multiplex DNAs detection and aptamer-substrate complexes sensing[J]. Analytical Chemistry, 2016, 89(1): 1002-1008.
[14] Liu S F, Zhang C X, Ming J J, et al. Amplified detection of DNA by an analyte-induced Y-shaped junction probe assembly followed with a nicking endonuclease-mediated
autocatalytic recycling process[J]. Chemical Communications,2013, 49(72): 7947-7949.
[15] Hu R, Liu T, Zhang X B, et al. Multicolor fluorescent biosensor for multiplexed detection of DNA[J]. Analytical Chemistry, 2014, 86(10): 5009-5016.
[16] He J A, Zhao F, Wu C L, et al. Development of a smart dynamic surface chemistry for surface plasmon resonance-based sensors for the detection of DNA molecules
[J]. Journal of Materials Chemistry B, 2013, 1(40): 5398-5402.
[17] Diao W, Tang M, Ding S J, et al. Highly sensitive surface plasmon resonance biosensor for the detection of HIV-related DNA based on dynamic and structural DNA nanodevices[
J]. Biosensors and Bioelectronics, 2018, 100:228-234.
[18] Zang Y, Lei J P, Zhang L, et al. In situ generation of electron acceptor for photoelectrochemical biosensing via hemin-mediated catalytic reaction[J]. Analytical Chemistry,
2014, 86(24): 12362-12368.
[19] Liu S L, Li C, Cheng J, et al. Selective photoelectrochemical detection of DNA with high-affinity metallointercalator and tin oxide nanoparticle electrode[J].Analytical Chemistry,
2006, 78(13): 4722-4726.
[20] Li C X, Wang H Y, Shen J, et al. Cyclometalated iridium complex-based label-free photoelectrochemical biosensor for DNA detection by hybridization chain reaction amplification[
J].Analytical Chemistry, 2015, 87(8): 4283-4291.
[21] Shankar K, Basham J I, Allam N K, et al. Recent advances in the use of TiO2 nanotube and nanowire arrays for oxidative photoelectrochemistry[J]. The Journal of Physical
Chemistry C, 2009, 113(16): 6327-6359.
[22] Tarwal N L, Patil P S. Enhanced photoelectrochemical performance of Ag-ZnO thin films synthesized by spray pyrolysis technique[J]. Electrochimica Acta, 2011, 56(18):
[23] Li D, Jia J L, Zheng T, et al. Construction and characterization of visible light active Pd nano-crystallite decorated and C-N-S-co-doped TiO2 nanosheet array photoelectrode
for enhanced photocatalytic degradation of acetylsalicylic acid[J]. Applied Catalysis B: Environmental, 2016, 188:259-271.
[24] Chang X W, Liu H L, Chen Q H, et al. Preparation of graphene film decorated TiO2 nano-tube array photoelectrode and its enhanced visible light photocatalytic mechanism[J]. Carbon, 2014, 66(3): 450-458.
[25] Lee Y L, Lo Y S. Highly efficient quantum-dot-sensitized solar cell based on co-sensitization of CdS/CdSe[J]. Advanced Functional Materials, 2009, 19(4): 604-609.
[26] Ge L, Sun X M, Hong Q, et al. Ratiometric catalyzed-assembly of nanocluster beacons: a nonenzymatic approach for amplified DNA detection[J]. ACS Applied Materials
and Interfaces, 2017, 9(37): 32089-32096.
[27] Hun X, Meng Y, Wang S S, et al. Mismatched catalytic hairpin assembly coupling hydroxylamine-o-sulfonic acid as oxide for DNA assay[J]. Sensors and Actuators B: Chemical,
2018, 254: 347-353.
[28] Wang Y, Gan N, Zhou Y, et al. Novel label-free and highthroughput microchip electrophoresis platform for multiplex antibiotic residues detection based on aptamer probes and target catalyzed hairpin assembly for signal amplification[J]. Biosensors and Bioelectronics, 2017, 97:100-106.
[29] Kivlehan F, Mavr佴F, Talini L, et al. Real-time electrochemical monitoring of isothermal helicase-dependent amplification of nucleic acids[J]. Analyst, 2011, 136(18):3635-3642.
[30] Ma F, Liu M, Tang B, et al. Sensitive quantification of microRNAs by isothermal helicase-dependent amplification[J]. Analytical Chemistry, 2017, 89(11): 6182-6187.
[31] Chen Y, Xu J, Su J, et al. In situ hybridization chain reaction amplification for universal and highly sensitive electrochemiluminescent detection ofDNA[J].AnalyticalChemistry,
2012, 84(18): 7750-7755.
[32] Ge Z L, Lin M H, Wang P, et al. Hybridization chain reaction amplification of microRNA detection with a tetrahedral DNA nanostructure-based electrochemical biosensor[J]. Analytical Chemistry, 2014, 86(4): 2124-2130.
[33] Yao G H, Liang R P, Yu X D, et al. Target-triggering multiplecycle amplification strategy for ultrasensitive detection of adenosine based on surface plasma resonance
techniques[J].Analytical Chemistry, 2014, 87(2): 929-936.
[34] Shen W, Deng H M, Gao Z Q. Gold nanoparticle-enabled real-time ligation chain reaction for ultrasensitive detection of DNA[J]. Journal of the American Chemical Society,
2012, 134(36): 14678-14681.
[35] Deng H, Xu Y, Liu Y H, et al. Gold nanoparticles with asymmetric polymerase chain reaction for colorimetric detection ofDNAsequence[J].AnalyticalChemistry, 2012,84(3): 1253-1258.
[36] Zhang K Y, Lv S Z, Lin Z Z, et al. Bio-bar-code-based photoelectrochemical immunoassay for sensitive detection of prostate-specific antigen using rolling circle amplification
and enzymatic biocatalytic precipitation[J]. Biosensors and Bioelectronics, 2018, 101: 159-166.
[37] Yan H, Xu Y C, Lu Y, et al. Reduced graphene oxidebased solid-phase extraction for the enrichment and detection of microRNA[J]. Analytical Chemistry, 2017, 89 (19): 10137-10140.
[38] Jiao M, Jie G F, Tan L, et al. AgNPs-3D nanostructure enhanced electrochemiluminescence of CdSe quantum dot coupled with strand displacement amplification for sensitive biosensing of DNA[J]. Analytica Chimica Acta,2017, 983: 166-172.
[39] Yin D, Tao Y Y, Tang L, et al. Cascade toehold-mediated strand displacement along with non-enzymatic target recycling amplification for the electrochemical determination
of the HIV-1 related gene[J]. Microchimica Acta,2017, 184(10): 3721-3728.
[40] Zheng A X, Li J, Wang J R, et al. Enzyme-free signal amplification in the DNAzyme sensor via target-catalyzed hairpin assembly[J]. Chemical Communications, 2012,48(25): 3112-3114.
[41] Li C X, Li Y X, Xu X, et al. Fast and quantitative differentiation of single-base mismatched DNA by initial reaction rate of catalytic hairpin assembly[J]. Biosensors and Bioelectronics, 2014, 60: 57-63.
[42] Ma C, Liu H Y, Zhang L N, et al. Multiplexed aptasensor for simultaneous detection of carcinoembryonic antigen and mucin-1 based on metal ion electrochemical labels
and Ru(NH3)63+ electronic wires[J]. Biosensors and Bioelectronics,2018, 99: 8-13.
[43] Tabrizi M A, Shamsipur M, Saber R, et al. A high sensitive visible light-driven photoelectrochemical aptasensor for shrimp allergen tropomyosin detection using graphitic
carbon nitride-TiO2 nanocomposite[J].Biosensors andBioelectronics,2017, 98: 113-118.
[44] Guo Q Q, Chen Y, Song Z P, et al. Label-free and enzyme-free sensitive fluorescent detection of human immunodeficiency virus deoxyribonucleic acid based on hybridization
chain reaction[J]. Analytica Chimica Acta, 2014,852: 244-249.
[45] Yang Y J, Huang J, Yang X H, et al. Gold nanoparticle based hairpin-locked-DNAzyme probe for amplified miRNA imaging in living cells[J]. Analytical Chemistry,2017, 89(11): 5850-5856.
[46] Quan K, Huang J, Yang X H, et al. An enzyme-free and amplified colorimetric detection strategy via target-aptamer binding triggered catalyzed hairpin assembly [J].Chemical Communications, 2015, 51(5): 937-940.
[47] Meng A Y, Zhu B C, Zhong B, et al. Direct Z-scheme TiO2/CdS hierarchical photocatalyst for enhanced photocatalytic H2-production activity[J]. Applied Surface Science,2017, 422: 518-527.
[48] Cui L, Li Y Y, Lu M F, et al. An ultrasensitive electrochemical biosensor for polynucleotide kinase assay based on gold nanoparticle-mediated lambda exonuclease cleavage-
induced signal amplification[J]. Biosensors and Bioelectronics,2018, 99: 1-7.
[49] Chen Y X, Huang K J, Lin F, et al. Ultrasensitive electrochemical sensing platform based on graphene wrapping SnO2 nanocorals and autonomous cascade DNA duplication
strategy[J]. Talanta, 2017, 175: 168-176.
[50] Wang ZH, Sun N, He Y, et al. DNA assembled gold nanoparticles polymeric network blocks modular highly sensitive electrochemical biosensors for protein kinase activity
analysis and inhibition[J]. Analytical Chemistry, 2014, 86(12): 6153-6159.
[51] Li S G, Zhu W, Xue Y C, et al. Construction of photoelectrochemical thrombin aptasensor via assembling multilayer of graphene-CdS nanocomposites[J]. Biosensors and
Bioelectronics, 2015, 64: 611-617.
[52] Zhang L, Sun Y, Liang Y Y, et al. Ag nanoclusters could efficiently quench the photoresponse of CdS quantum dots for novel energy transfer-based photoelectrochemical
bioanalysis[J]. Biosensors and Bioelectronics, 2016, 85:930-934.
[53] Wang G L, Liu K L, Shu J X, et al. A novel photoelectrochemical sensor based on photocathode of PbS quantum dots utilizing catalase mimetics of bio-bar-coded platinum
nanoparticles/G-quadruplex/hemin for signal amplification[J]. Biosensors and Bioelectronics, 2015, 69: 106-112.
[54] Gao Z Q, Tansil N C. An ultrasensitive photoelectrochemical nucleic acid biosensor[J]. Nucleic Acids Research,2005, 33(13): e123.
[55] Xiong E, Yan X, Zhang X H, et al. A new photoelectrochemical biosensor for ultrasensitive determination of nucleic acid based on three-stage cascade signal amplification
strategy[J]. Analyst, 2018, 143: 2799-2806.



To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.