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Article | Open Access

Real time PCR based on Fluorescent Quenching of Mercaptoacetic Acid-Modified CdTe Quantum Dots for Ultrasensitive Specific Detection of Nucleic Acids

Daxiang Cui1( ), Qing Li1, Peng Huang1, Kan Wang1, Yifei Kong1, Hong Zhang2, Xiaogang You1, Rong He1, Hua Song1, Jingping Wang2, Chenchen Bao1, Toru Asahi2, Feng Gao1, Tetsuya Osaka2
Department of Bio-Nano Science and Engineering, National Key Laboratory of Micro/Nano Fabrication Technology, Thin Film Key Laboratory of Education Department, Institute of Micro-Nano Science and Technology, Shanghai JiaoTong University, 200030, Shanghai, P. R. China
Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 513 Wasedatsurumaki-cho, Shinjuku-ku,Tokyo 162-0041, Japan
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Abstract

A ultra-sensitive, highly specific, real-time polymerase chain reaction system based on mercaptoacetic acid-modified CdTe nanocrystals(mQDs) is reported. With the addition of 3 nm mQDs into the PCR reagent, the photoluminescent(PL) intensities of mQDs decreased gradually as the DNA templates and PCR cycles increased, in an approximate negative linear relation to the DNA concentration logarithm or cycles, the PL peaks exhibited red-shifts synchronously. Mg2+ ions decreased the PL intensity of mQDs in a dose-dependent means, and Taq DNA polymerase enhanced the PL intensity of mQDs in a dose-dependent means. Real-time PCR based on mQDs showed an increased sensitivity at least 103 fold higher than that based on SYBR Green I. The specificity of PCR was enhanced in the PCR reagent with less than 1.33mg/mL mQDs. The potential mechanism is also discussed. This novel PCR system based on mQDs has great potential in applications such as ultra-sensitive specific DNA or RNA detection, dynamic molecular imaging, and photoelectric biosensors.

Keywords

SpecificityPhotoluminescenceSensitivityMercaptoacetic acid-modified CdTe quantum dotPolymerase chain reaction

References

[1]

Jeong H, Chang AM, Melloch MR, The Kondo effect in an artificial quantum dot molecule. Science 2001; 293: 2221-2223.doi: 10.1126/science.1063182
Crossref Google Scholar
[2]

Cui D, Han Y, Li Z, Song H, Wang K, He R, et al. Fluorescent magnetic nanoprobes for in vivo targeted imaging and hyperthermia therapy of prostatecancer. Nano Biomed Eng 2009; 1(1): 94-112.
Crossref Google Scholar
[3]

Li Z, Huang P, He R, Lin J, Yang S, Zhang X, et al. Aptamer-conjugated dendrimer-modified quantum dots for cancer cell targeting and imaging. Mater. Lett. 2010; 64: 375–378.doi: 10.1016/j.matlet.2009.11.022
Crossref Google Scholar
[4]

Lu, W, Ji Z Q, Pfeiffer, L, West, K W, Rimberg, A J. Realtime detection of electron tunnelling in a quantum dot. Nature 2003; 423: 422-425.doi: 10.1038/nature01642
Crossref Google Scholar
[5]

Kaul, Z, Yaguchi T, Kaul SC Hirano, T Wadhwa R, Taira K. Mortalin imaging in normal and cancer cells with quantum dot immuno-conjugates. Cell Res. 2003; 13: 503-507.doi: 10.1038/sj.cr.7290194
Crossref Google Scholar
[6]

Medintz IL, Uyeda HT, Goldman ER, Mattoussi H. Quantum dot bioconjugates for imaging, labelling and sensing. Nature Mater. 2005; 4: 435-446.doi: 10.1038/nmat1390
Crossref Google Scholar
[7]

Yang H, Guo Q, He R, Li D, Zhang XQ, Bao CC, Hu HY, Cui DX. A quick and parallel analytical method based on quantum dots labeling for ToRCH-related antibodies. Nanoscale Res. Lett. 2009; 4: 1469-1474.doi: 10.1007/s11671-009-9422-7
Crossref Google Scholar
[8]

Cui DX, Pan BF, Zhang H, et al. Self-assembly of CNTs and Quantum dots for ultrasensitive DNA and antigen detction.Analytical Chemsitry, 2008, 80(21)7996-8001.doi: 10.1021/ac800992m
Crossref Google Scholar
[9]

Han M, Gao X, Su J Z, Nie SM. Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules. Nature Biotechnol. 2001; 19: 631-635.doi: 10.1038/90228
Crossref Google Scholar
[10]

Jares-Erijman E A, Jovin T M. FRET imaging. Nature Biotechnology 2003; 21: 1387-1395.doi: 10.1038/nbt896
Crossref Google Scholar
[11]

Huang X Y, Li L, Qian H F, Dong C Q, Ren C J. A resonance energy transfer between chemiluminescent donors and luminescent quantum-dots as acceptors (CRET). Angew. Chem. Int. Ed. 2006; 45: 5140-5143. doi: 10.1002/anie.200601196
Crossref Google Scholar
[12]

Alivisato P. The use of nanocrystals in biological detection. Nature Biotechnol. 2004; 22: 47-52.doi: 10.1038/nbt927
Crossref Google Scholar
[13]

Dong CQ, Qian HF, Fang NH, Ren JC. Study of fluorescence quenching and dialysis process of CdTe quantum dots, using ensemble techniques and fluorescence correlation spectroscopy. J. Phys. Chem. B. 2006; 110: 11069-11075.doi: 10.1021/jp060279r
Crossref Google Scholar
[14]

Ullis KB, Faloona FA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987; 155: 335-350.doi: 10.1016/0076-6879(87)55023-6
Crossref Google Scholar
[15]

Kubista M, Andrade JM, Bengtsson M, Forootan A, Jonak J, Lind K, Sindelka R, Sjoback R, Sjogreen B, Strombom L, Stahlberg A, Zoric N. The real-time polymerase chain reaction. Mol. Aspects Med. 2006; 27: 95-125.doi: 10.1016/j.mam.2005.12.007
Crossref Google Scholar
[16]

Chen F, Binder B, Hodson RE. Flow cytometric detection of specific gene expression in prokaryotic cells using in situ RT-PCR. FEMS Microbiol. 2000; 184: 291-295. doi: 10.1111/j.1574-6968.2000.tb09029.x
Crossref Google Scholar
[17]

Higuchi R, Dollinger G, Walsh PS, Griffith R. Simultaneous amplification and detection of specific DNAsequences. Biotechnology 1992; 10:423-417.
Crossref Google Scholar
[18]

Lagally E T, Simpson PC, Mathies R A. Monolithic integrated microfluidic DNA amplification and capillary electrophoresis analysis system. Sensors Actuators B 2000; 63: 38-146.doi: 10.1016/S0925-4005(00)00350-6
Crossref Google Scholar
[19]

Cui D, Tian F R, Kong Y, Igor T, Gao H. Effects of Single wall carbon nnaotubes on polymerase chain reaction. Nanotechnology 2004;15:154-157.doi: 10.1088/0957-4484/15/1/030
Crossref Google Scholar
[20]

Shen CC, Yang W J, Ji Q L, Maki H, Dong A J, Zhang Z Z. NanoPCR observation: different levels of DNA replication fidelity in nanoparticle enhanced polymerase chain reactions. Nanotechnology 2009; 20: 455103. doi: 10.1088/0957-4484/20/45/455103
Crossref Google Scholar
[21]

Li Q, He R, Gao F, Cui, D. The Effect s of mercaptoacetic acid modified CdTe quantum dot s on specificity of polymerase chain reaction. Journal of Shanghai Jiaotong university 2008; 5: 693-696.
Google Scholar
[22]

Li H, Huang J, Lv J, An H, Zhang X, Zhang Z, Fan C, Hu J. Nanoparticle PCR: Nanogold-assisted PCR with enhanced specificity. Angew. Chem. Int. Ed. 2005; 44: 5100-5103. doi: 10.1002/anie.200500403
Crossref Google Scholar
[23]

Li M, Lin YC, Wu C C, Liu H S. Enhancing the efficiency of a PCR using gold nanoparticles. Nucleic Acids Research. 2005; 33: e184.doi: 10.1093/nar/gni183
Crossref Google Scholar
[24]

Yang W C, Mi L, Cao X Y, Zhang X D, Fan C H, Hu J. Evaluation of gold nanoparticles as the additive in real-time polymerase chain reaction with SYBR Green I dye. Nanotechnology 2008; 19: 255101.doi: 10.1088/0957-4484/19/25/255101
Crossref Google Scholar
[25]

Cui D, Jin G, Gao T W, Sun T, Tian F, Estrada G G, Gao H, Sarai A. Characterization of BRCAA1 and its novel epitope identification. Cancer Epidemiol Biomarkers Prev. 2004; 13: 1136-1145.
Crossref Google Scholar
[26]

He R, You X, Shao J, Gao F, Pan B, Cui D. Core/shell fluorescent magnetic silica-coated composite nanoparticles for bioconjugation. Nanotechnology.2007; 18 (31):315601. doi: 10.1088/0957-4484/18/31/315601
Crossref Google Scholar
[27]

Mamedova N N, Kotov NA. Albumin-CdTe Nanoparticle Bioconjugates: Preparation, Structure, and Interunit Energy Transfer with Antenna effect.Nano Lett. 2001; 1: 281-286. doi: 10.1021/nl015519n
Crossref Google Scholar
[28]

Asuri P, Bale SS, Karajanagi S S, Kane R S. The protein-nanomaterial interface. Current opinion in biotechnology 2006; 17: 562-568.doi: 10.1016/j.copbio.2006.09.002
Crossref Google Scholar
[29]

Ponchel F, Toomes C, Bransfield K, Leong F T, Douglas S H, Field S L, Bell S M, Combaret V, Puisieux A, Mighell A J et al. Real time PCR based on SYBR-Green I fluorescence: an alternative to the TaqMan assay for a relative quantification of gene rearrangements, gene amplifications, and micro gene delections. BMC Biotechnol. 2003; 3:18-20.doi: 10.1186/1472-6750-3-18
Crossref Google Scholar
[30]

Wang Y M, Zhu W, Levy D E. Nuclear and cytoplasmic mRNA quantification by SYBR green based real-time RTPCR. Methods 2006; 39: 356-362.doi: 10.1016/j.ymeth.2006.06.010
Crossref Google Scholar
[31]

Ji XQ, Sato H, Tanaka H, Konishi Y, Fujimoto T, Takahashi O, Tanaka T. Real-time quantitative RT-PCR detection of disseminated endometrial tumor cells in peripheral blood and lymph nodes using the LightCycler System. Gynecol Oncol. 2006; 100: 355-60. doi: 10.1016/j.ygyno.2005.08.041
Crossref Google Scholar
[32]

Wright P A, Wynford-Thomas D. The polymerase chain reaction: miracle or mirage? A critical review of its uses and limitations in diagnosis and research. J Pathol.1990; 162: 99-117.doi: 10.1002/path.1711620203
Crossref Google Scholar
[33]

Talapin D V, Gaponik N, Borchert H, Rogach A L, Haase M, Weller H. Etching of colloidal InP nanocrystals with fluorides: Photochemical nature of the process resulting in high photoluminescence efficiency. J. Phys. Chem. B. 2002; 106: 7177-7180.doi: 10.1021/jp026380n
Crossref Google Scholar
[34]

Helot A, Favero I, Cassabois G, Voisin C, Delalande C, Roussignol Ph, Ferreira R, Gerard J M. Unconventional motional narrowing in the optical spectrum of a semiconductor quantum dot. Nature Physics 2006; 2: 759-764.doi: 10.1038/nphys433
Crossref Google Scholar
[35]

Wang P. Nanoscale bio-catalyst systems. Current Opinion in biotechnology 2006; 17: 74-579.doi: 10.1016/j.copbio.2006.10.009
Crossref Google Scholar
[36]

Bigion T P, Lin X M, Nguyen T T, Corwin E I, Witten T A, Jaeger H M. Kinetically driven self assembly of highly ordered nnaoparticle monolayers. Nature Materials 2006; 5: 265-270.doi: 10.1038/nmat1611
Crossref Google Scholar
[37]

Eijt S W H, Veen A V, Schut H, Mijnarends P E, Denison A B, Barbiellini B, Bansil A A. Study of colloid quantumdot surfaces using an innovative thin-film positron 2D-ACAR method. Nature Materials 2006; 5: 23-26. doi: 10.1038/nmat1550
Crossref Google Scholar
[38]

Mitchell G P, Mirkin C A, Lrtsinger R L. Programmed assembly of DNA functionalized quantum dots J. Am.Chem.soc. 1999; 121: 8122-8124.doi: 10.1021/ja991662v
Crossref Google Scholar
[39]

Gaponik N, Talapin D V, Rogach A L, Hoppe K, Shevchenko E V, Kornowski A, Eychmuller A, Weller H. Thiol-capping of CdTe nanocrystals: An alternative to organometallic synthetic routes. J. Physics Chem. B 2002; 106: 7177-7185.doi: 10.1021/jp025541k
Crossref Google Scholar
[40]

Kenworthy A K. Imaging protein-protein interactions using fluorescence resonance energy transfer microscpy. Methods 2001; 24: 289-296.doi: 10.1006/meth.2001.1189
Crossref Google Scholar
[41]

Perales C, Cava F, Meijer W J J, Berenguer J. Enhancement of DNA, cDNA synthesis and fidelity at high temperatures by a dimeric single-stranded DNA-binding protein. Nucleic Acids Res.2003; 31: 6473-6480.doi: 10.1093/nar/gkg865
Crossref Google Scholar
[42]

Lee S H, Taguchi H, Yoshimura E, Minagawa E, Kaminogawa S, Ohta T, Matsuzawa H. Carboxypeptidase Taq, a thermostable zinc enzyme, from Thermus aquaticus YT-1: molecular cloning, sequencing, and expression of the encoding gene in Escherichia coli. J. Biosci. Biotechnol. Biochem. 1994; 58: 1490-1495.doi: 10.1271/bbb.58.1490
Crossref Google Scholar
[43]

Jiao G S, Thoresen L H, Kim T G, Haaland W C, Gao F, Topp M R, Hochstrasser R M, Metzker M L, Burgess K. Syntheses, photophysical properties, and application of through-bond energy-transfer cassettes for biotechnology. Chemistry -- A European Journal 2006; 12: 7816-7826. doi: 10.1002/chem.200600197
Crossref Google Scholar
[44]

Chou Q. Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res. 1992; 20: 1717-1723. doi: 10.1093/nar/20.7.1717
Crossref Google Scholar
[45]

Chou Q. Minimizing deletion mutagenesis artifact during Taq DNA polymerase PCR by E. coli SSB. Nucleic Acids Res. 1992; 20: 4371.doi:10.1093/nar/20.16.4371
Crossref Google Scholar
Nano Biomedicine and Engineering
Volume 2 Issue 1,
March 2010
Pages 45-55
DOI: 10.5101/nbe.v2i1.p45-55
Cite this article:
Cui D, Li Q, Huang P, et al. Real time PCR based on Fluorescent Quenching of Mercaptoacetic Acid-Modified CdTe Quantum Dots for Ultrasensitive Specific Detection of Nucleic Acids. Nano Biomedicine and Engineering, 2010, 2(1): 45-55. https://doi.org/10.5101/nbe.v2i1.p45-55

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Received: 15 January 2010
Accepted: 15 February 2010
Published: 05 March 2010
© 2010 D. Cui et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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