Abstract
Prostate-specific antigen (PSA) is the only biomarker for the diagnosis of prostate cancer. So the PSA screening test is very important due to the high occurrence of prostate cancer in men. In this work, a label-free fluorescent method was developed based on terminal deoxynucleotidyl transferase (TdT) and G–quadruplex–thioflavin T complex for detecting PSA. In the absence of PSA, the PSA aptamer can be used as the primer for TdT extension reactions, resulting in the formation of G-quadruplexes and generation of strong fluorescent signals. After the addition of PSA, the PSA–aptamer complex prevented the TdT extension reaction due to steric hindrance, thus resulting in a poor fluorescent signal. The assay showed a wide linear range (0.1 to 80 pg/mL) and a detection limit of 0.086 pg/mL (S/N = 3). It also has good specificity for PSA determination and gives satisfactory results when applied to biological samples. Conceivably, its merits such as good selectivity and high sensitivity indicate that the proposed method has a promising application potential in the clinical diagnosis and treatment of prostate cancer.
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References
Díaz-Fernández A, Miranda-Castro R, de-los-Santos-Álvarez N, Rodríguez EF, Lobo-Castañón MJ. Focusing aptamer selection on the glycan structure of prostate-specific antigen: toward more specific detection of prostate cancer. Biosens Bioelectron 2019;128:83–90.
Kong RM, Ding L, Wang ZJ, You JM, Qu FL. A novel aptamer-functionalized MoS2 nanosheet fluorescent biosensor for sensitive detection of prostate specific antigen. Anal Bioanal Chem. 2015;407:369–77.
Chen Y, Yuan PX, Wang AJ, Luo XL, Xue YD, Zhang L, et al. A novel electrochemical immunosensor for highly sensitive detection of prostate-specific antigen using 3D open-structured PtCu nanoframes for signal amplification. Biosens Bioelectron. 2019;126:187–92.
Wei B, Mao K, Liu N, Zhang M, Yang ZG. Graphene nanocomposites modified electrochemical aptamer sensor for rapid and highly sensitive detection of prostate specific antigen. Biosens Bioelectron. 2018;121:41–6.
Cao JT, Yang JJ, Zhao LZ, Wang YL, Wang H, Liu YM, et al. Graphene oxide@gold nanorods-based multiple-assisted electrochemiluminescence signal amplification strategy for sensitive detection of prostate specific antigen. Biosens Bioelectron. 2018;99:92–8.
Zhou Q, Lin YX, Zhang KY, Li MJ, Tang DP. Reduced graphene oxide/BiFeO3 nanohybrids-based signal-on photoelectrochemical sensing system for prostate specific antigen detection coupling with magnetic microfluidic device. Biosens Bioelectron. 2018;101:146–52.
Zhang KY, Lv SZ, Lin ZZ, Li MJ, Tang DP. Bio-bar-code-based photoelectrochemical immunoassay for sensitive detection of prostate-specific antigen using rolling circle amplification and enzymatic biocatalytic precipitation. Biosens Bioelectron. 2018;101:159–66.
Barbosa AI, Wichers JH, van Amerongen A, Reis NM. Towards one-step quantitation of prostate-specific antigen (PSA) in microfluidic devices: feasibility of optical detection with nanoparticle labels. Bionanoscience. 2017;7:718–26.
Chen Y, Guo XY, Liu W, Zhang L. Paper-based fluorometric immunodevice with quantum-dot labeled antibodies for simultaneous detection of carcinoembryonic antigen and prostate specific antigen. Microchim Acta 2019;186:112.
Zhang LJ, Luo ZB, Zeng RJ, Zhou Q, Tang DP. All-solid-state metal-mediated Z-scheme photoelectrochemical immunoassay with enhanced photoexcited charge-separation for monitoring of prostate-specific antigen. Biosens Bioelectron. 2019;134:1–7.
Lv SZ, Zhang KY, Zeng YY, Tang DP. Double photosystems-based ‘Z-Scheme’ photoelectrochemical sensing mode for ultrasensitive detection of disease biomarker accompanying three-dimensional DNA walker. Anal Chem. 2018;90:7086–93.
Tian CY, Wang L, Luan F, Zhuang XM. An electrochemiluminescence sensor for the detection of prostate protein antigen based on the graphene quantum dots infilled TiO2 nanotube arrays. Talanta. 2019;191:103–8.
Gao ZQ, Lv SZ, Xu MD, Tang DP. High-index {hk0} faceted platinum concave nanocubes with enhanced peroxidase-like activity for an ultrasensitive colorimetric immunoassay of the human prostate-specific antigen. Analyst. 2017;142:911–7.
Shao K, Wang B, Nie A, Ye SY, Ma J, Li ZG, et al. Target-triggered signal-on ratiometric electrochemiluminescence sensing of PSA based on MOF/au/ G-quadruplex. Biosens Bioelectron. 2018;118:160–6.
Yazdani Z, Yadegari H, Heli H. A molecularly imprinted electrochemical nanobiosensor for prostate specific antigen determination. Anal Biochem. 2019;566:116–25.
Li YY, Khan MS, Tian LH, Liu L, Hu LH, Fan DW, et al. An ultrasensitive electrochemical immunosensor for the detection of prostate-specific antigen based on conductivity nanocomposite with halloysite nanotubes. Anal Bioanal Chem. 2017;409:3245–51.
Damborska D, Bertok T, Dosekova E, Holazova A, Lorencova L, Kasak P, et al. Nanomaterial-based biosensors for detection of prostate specific antigen. Microchim Acta. 2017;184:3049–67.
Fang BY, An J, Liu B, Zhao YD. Hybridization induced fluorescence enhanced DNA-ag nanocluster/aptamer probe for detection of prostate-specific antigen. Colloids Surf B Biointerfaces. 2019;175:358–64.
Wu KF. Ma CB, Zhao H, He HL. Chen HC Label-free G-quadruplex aptamer fluorescence assay for ochratoxin A using a thioflavin T probe Toxins. 2018;10:198.
Tang ZW, Chen HT, He HL, Ma CB. Assays for alkaline phosphatase activity: progress and prospects. Trends Anal Chem. 2019;113:32–43.
Wu KF, Ma CB, Zhao H, Chen MJ, Deng ZY. Sensitive aptamer-based fluorescene assay for ochratoxin a based on RNase H signal amplification. Food Chem. 2019;277:273–8.
Zhao H, Xiang XY, Chen MJ, Ma CB. Aptamer-based fluorometric ochratoxin a assay based on photoinduced electron transfer. Toxins. 2019;11:65.
Chi BZ, Liang RP, Yuan YH, Zhang L, Li ZM, Qiu JD. Luminescence determination of microRNAs based on the use of terbium (III) sensitized with an enzyme-activated guanine-rich nucleotide. Microchim Acta. 2018;185:280.
Tang ZW, Zhang HF, Ma CB, Gu P, Zhang GH, Wu KF, et al. Colorimetric determination of the activity of alkaline phosphatase based on the use of cu (II)-modulated G-quadruplex-based DNAzymes. Microchim Acta. 2018;185:109.
Zhao H, Ma CB, Chen MJ. A novel fluorometric method for inorganic pyrophosphatase detection based on G-quadruplex-thioflavin T. Mol Cell Probes. 2019;43:29–33.
Liu Z, Lei S, Zou LN, Li GP, Xu LL, Ye BX. A label-free and double recognition–amplification novel strategy for sensitive and accurate carcinoembryonic antigen assay. Biosens Bioelectron. 2019;131:113–8.
Ma CB, Wu KF, Zhao H, Liu HS, Wang KM, Xia K. Fluorometric aptasensor of ochratoxin a based on the use of graphene oxide and RNase H-aided amplification. Microchim Acta. 2018;185:347.
Gu P, Zhang GH, Deng ZY, Tang ZW, Zhang HF, Khusbu FY, Ket al. A novel label-free colorimetric detection of L-histidine using Cu2+-modulated G-quadruplex-based DNAzymes. Spectrochim Acta A Mol Biomol Spectrosc 2018;203:195–200.
Mohanty J, Barooah N, Dhamodharan V, Harikrishna S, Pradeepkumar PI, Bhasikuttan AC. Thioflavin T as an efficient inducer and selective fluorescent sensor for the human telomeric G-quadruplex DNA. J Am Chem Soc. 2013;135:367–76.
Khusbu FY, Zhou X, Chen HC, Ma CB, Wang KM. Thioflavin T as a fluorescence probe for biosensing applications. Trends Anal Chem. 2018;109:1–18.
Xu HY, Geng FH, Jiang XY, Shao CY, Wang YX, Wang KF, et al. Design of metal-ion-triggered assembly of label-free split G-quadruplex/duplex DNA for turn-on detection of Hg2+ in fetal calf serum. Sens Actuators B Chem. 2018;255:1024–30.
Liu SG, Luo D, Han L, Li NB, Luo HQ. A hybrid material composed of guanine-rich single stranded DNA and cobalt (III) oxyhydroxide (CoOOH) nanosheets as a fluorescent probe for ascorbic acid via formation of a complex between G-quadruplex and thioflavin T. Microchim Acta. 2019;186:156.
Chen MJ, Deng ZY, Ma CB, Zhao H, Wu KF, Wang KM. A sensitive fluorescence method for the detection of streptavidin based on target-induced DNA machine amplification. Anal Methods. 2018;10:1870–4.
Li YN, Wang JY, Zhang B, He Y, Wang JP, Wang S. A rapid fluorometric method for determination of aflatoxin B1 in plant-derived food by using a thioflavin T-based aptasensor. Microchim Acta. 2019;186:214.
Tang XX, Wu KF, Zhao H, Chen MJ, Ma CB. A label-free fluorescent assay for the rapid and sensitive detection of adenosine deaminase activity and inhibition. Sensors. 2018;18:2441.
Wu KF, Ma CB, Deng ZY, Fang N, Tang ZW, Zhu XX, et al. Label-free and nicking enzyme-assisted fluorescence signal amplification for RNase H determination based on a G-quadruplexe/thioflavin T complex. Talanta. 2018;182:142–7.
Liu Z, Li W, Nie Z, Peng F, Huang Y, Yao S. Randomly arrayed G-quadruplexes for label-free and real-time assay of enzyme activity. Chem Comm. 2014;50:6875–8.
Cao Y, Wang ZH, Cao JP, Mao XX, Chen GF, Zhao J. A general protein aptasensing strategy based on untemplated nucleic acid elongation and the use of fluorescent copper nanoparticles: application to the detection of thrombin and the vascular endothelial growth factor. Microchim Acta. 2017;184:3697–704.
You PY, Li FC, Liu MH, Chan YH. Colorimetric and fluorescent dual-mode immunoassay based on plasmon-enhanced fluorescence of polymer dots for detection of PSA in whole blood. ACS Appl Mater Interfaces. 2019;11:9841–9.
Hwang DG, Chae YM, Choi N, Cho IJ, Kang JY, Lee SH. Label-free detection of prostate specific antigen (PSA) using a bridge-shaped PZT resonator. Microsyst Technol. 2017;23:1207–14.
Cai GN, Yu ZZ, Ren RG, Tang DP. Exciton-plasmon interaction between AuNPs/graphene nanohybrids and CdS quantum dots/TiO2 for photoelectrochemical aptasensing of prostate-specific antigen. ACS Sens. 2018;3:632–9.
Zhou Q, Lin YX, Shu J, Zhang KY, Yu ZZ, Tang DP. Reduced graphene oxide-functionalized FeOOH for signal-on photoelectrochemical sensing of prostate-specific antigen with bioresponsive controlled release system. Biosens Bioelectron. 2017;98:15–21.
Zhang KY, Lv SZ, Lin ZZ, Tang DP. CdS:Mn quantum dot-functionalized g-C3N4 nanohybrids as signal-generation tags for photoelectrochemical immunoassay of prostate specific antigen coupling DNAzyme concatamer with enzymatic biocatalytic precipitation. Biosens Bioelectron. 2017;95:34–40.
Sun YJ, Wang CY, Zhang H, Zhang YL, Zhang GJ. A non-enzymatic and label-free fluorescence bioassay for ultrasensitive detection of PSA. Molecules. 2019;24:831.
Acknowledgments
This work was supported by State Key Laboratory of Chemo/ Biosensing and Chemometrics, Hunan University (2017006), The Research Innovation Program for Graduates of Central South University (2018zzts384, 2018zzts399, 2019zzts453).
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Chen, M., Ma, C., Yan, Y. et al. A label-free fluorescence method based on terminal deoxynucleotidyl transferase and thioflavin T for detecting prostate-specific antigen. Anal Bioanal Chem 411, 5779–5784 (2019). https://doi.org/10.1007/s00216-019-01958-0
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DOI: https://doi.org/10.1007/s00216-019-01958-0