基于凝血酶与腺苷核酸适配体功能化纳米探针识别的分析方法研究
|
摘要
纳米探针技术和核酸适配体识别是近年来迅速发展的新型探针技术和分子识别模式,在生物分析、药物检测、食品安全检测、临床检验、环境分析等方面表现出了巨大的应用潜力。发光分析技术因为具有灵敏度高、线性范围宽等优点,近年来也越来越受重视。论文以凝血酶和腺苷特异结合核酸适配体功能化金纳米粒子构建新型纳米探针,结合化学发光检测技术和金染、银染信号放大技术,以及均相共振光散射技术,建立了适用于凝血酶和腺苷高灵敏检测的方法体系。相关分析技术的发展,对于丰富纳米探针和发光分析技术理论,拓宽其应用具有积极意义。
首先,将核酸适配体功能化金纳米粒子与核酸适配体识别技术相结合,以凝血酶为模式分析物,结合纳米金溶出流动注射化学发光分析技术,建立基于核酸适配体功能化金纳米粒子识别的凝血酶化学发光新型检测方法。方法检测限达0.3×10-10 g/mL。
其次,在前面研究的基础上,以凝血酶蛋白为模式分析物,采用凝血酶特异核酸适配体识别凝血酶,并通过特异识别反应将凝血酶核酸适配体功能化纳米金探针组装到固相表面,进一步在纳米金颗粒表面进行金的选择性沉积,实现第一次信号放大,然后采用溶出化学发光检测实现第二次信号放大,使得该生物传感系统的灵敏度大大提高。
接着,针对金染化学溶出相对费时的问题,第四章继续以凝血酶蛋白为模式分析物,采用凝血酶特异核酸适配体识别凝血酶,并通过特异识别反应将凝血酶核酸适配体功能化纳米金探针组装到固相表面,进一步在纳米金颗粒表面进行银的选择性沉积,实现第一次原位信号放大,然后采用溶出化学发光检测实现第二次信号放大,建立一种适用于凝血酶超痕量检测的新型方法体系。方法检测限达0.3×10-12 g/mL。
最后,将纳米探针技术、核酸适配体识别与共振光散射检测技术联用,通过均相体系中腺苷与其特异结合核酸适配体识别反应,使形成网络聚集结构的金纳米结构解聚,可引起共振光散射信号强烈变化。基于此,建立了核酸适配体识别的均相检测腺苷的新方法。
Nano-probe technique and aptamer recognition are novel probe technique and moleculars recognition mode developing rapidly in recent years, which have presented tremendously potential in the application of biological analysis, drug testing, food safety detection, clinical verify, environmental analysis and the others. Luminescnect analysis with the merits of high sensitivity and wide linear range has also attracted more and more attentions recently. This paper means to construct novel nano-probes using thrombin and adenosine binding with their specific aptamer-functionalized Au nanoparticles, combining with chemiluminescence detecting technology, the Au and Ag staining signal amplification technology and resonance light scattering technology, establish methods systems suit for highly sensitive detection of thrombin and adenosine. In the meanwhile, electrochemi -luminescece analysis was combined with capillary electrophoresis separation to establish a rapidly and sensitively analytical method for clarithromycin determination in biological fluids. All these would be of importance for enriching the theory and accelerating the applications of nano-probe and luminescent analysis technique.
Firstly, combining the aptamer-functionalized Au nanoparticles with aptamer recognition technology, using thrombin as the model analyte, a novel analytical method for thrombin was developed coupling with flow-injection nanogold stripping chemiluminescence analysis technique.The dectiong limit of this method is 0.3×10-10 g/mL.
Secondly, based on above reseach, using thrombin as model analyte, introducing its specific aptamer recognise thrombin, thrombin-specific aptamer functionalized Au nano-probe were assembled to the solid matrix surface. Then a selective gold deposition procedure on gold nanoparticles surface was carried out to realize the first signal amplification. Stripping chemiluminescence detection was conducted to further amplify the detection signals, making the sensitivity of the biological sensing system enhanced greatly.
Thirdly, considering the problem of time consumption in gold stain, similar to gold staining based stripping CL detection, Chapter IV selectively deposite silver on the surface of nanogold for the first in-situ signal amplification, and then bond with the stripping chemiluminescence detection to carry out the second signal amplification, established a novel ultra-trace detection method for thrombin. The dectiong limit of this method is 0.3×10-12 g/mL.
Finally, coupling nano-probe technology, aptamer recognition with resonance light scattering analysis, through recognising reaction between adenosine and its specific aptamer in homogeneous matrix, causing the deaggregation of the aggregated network structure of adenosine specific aptamer functionalized nanogold structure, a sharply change on the resonance light scattering signal can be obtained. Based on it, a novel homogeneous aptamer recognizing based method for adenosine detection was established.
首先,将核酸适配体功能化金纳米粒子与核酸适配体识别技术相结合,以凝血酶为模式分析物,结合纳米金溶出流动注射化学发光分析技术,建立基于核酸适配体功能化金纳米粒子识别的凝血酶化学发光新型检测方法。方法检测限达0.3×10-10 g/mL。
其次,在前面研究的基础上,以凝血酶蛋白为模式分析物,采用凝血酶特异核酸适配体识别凝血酶,并通过特异识别反应将凝血酶核酸适配体功能化纳米金探针组装到固相表面,进一步在纳米金颗粒表面进行金的选择性沉积,实现第一次信号放大,然后采用溶出化学发光检测实现第二次信号放大,使得该生物传感系统的灵敏度大大提高。
接着,针对金染化学溶出相对费时的问题,第四章继续以凝血酶蛋白为模式分析物,采用凝血酶特异核酸适配体识别凝血酶,并通过特异识别反应将凝血酶核酸适配体功能化纳米金探针组装到固相表面,进一步在纳米金颗粒表面进行银的选择性沉积,实现第一次原位信号放大,然后采用溶出化学发光检测实现第二次信号放大,建立一种适用于凝血酶超痕量检测的新型方法体系。方法检测限达0.3×10-12 g/mL。
最后,将纳米探针技术、核酸适配体识别与共振光散射检测技术联用,通过均相体系中腺苷与其特异结合核酸适配体识别反应,使形成网络聚集结构的金纳米结构解聚,可引起共振光散射信号强烈变化。基于此,建立了核酸适配体识别的均相检测腺苷的新方法。
Nano-probe technique and aptamer recognition are novel probe technique and moleculars recognition mode developing rapidly in recent years, which have presented tremendously potential in the application of biological analysis, drug testing, food safety detection, clinical verify, environmental analysis and the others. Luminescnect analysis with the merits of high sensitivity and wide linear range has also attracted more and more attentions recently. This paper means to construct novel nano-probes using thrombin and adenosine binding with their specific aptamer-functionalized Au nanoparticles, combining with chemiluminescence detecting technology, the Au and Ag staining signal amplification technology and resonance light scattering technology, establish methods systems suit for highly sensitive detection of thrombin and adenosine. In the meanwhile, electrochemi -luminescece analysis was combined with capillary electrophoresis separation to establish a rapidly and sensitively analytical method for clarithromycin determination in biological fluids. All these would be of importance for enriching the theory and accelerating the applications of nano-probe and luminescent analysis technique.
Firstly, combining the aptamer-functionalized Au nanoparticles with aptamer recognition technology, using thrombin as the model analyte, a novel analytical method for thrombin was developed coupling with flow-injection nanogold stripping chemiluminescence analysis technique.The dectiong limit of this method is 0.3×10-10 g/mL.
Secondly, based on above reseach, using thrombin as model analyte, introducing its specific aptamer recognise thrombin, thrombin-specific aptamer functionalized Au nano-probe were assembled to the solid matrix surface. Then a selective gold deposition procedure on gold nanoparticles surface was carried out to realize the first signal amplification. Stripping chemiluminescence detection was conducted to further amplify the detection signals, making the sensitivity of the biological sensing system enhanced greatly.
Thirdly, considering the problem of time consumption in gold stain, similar to gold staining based stripping CL detection, Chapter IV selectively deposite silver on the surface of nanogold for the first in-situ signal amplification, and then bond with the stripping chemiluminescence detection to carry out the second signal amplification, established a novel ultra-trace detection method for thrombin. The dectiong limit of this method is 0.3×10-12 g/mL.
Finally, coupling nano-probe technology, aptamer recognition with resonance light scattering analysis, through recognising reaction between adenosine and its specific aptamer in homogeneous matrix, causing the deaggregation of the aggregated network structure of adenosine specific aptamer functionalized nanogold structure, a sharply change on the resonance light scattering signal can be obtained. Based on it, a novel homogeneous aptamer recognizing based method for adenosine detection was established.
引文
1.王楠,徐淑坤,王文星.纳米金生物探针及其应用[J].化学进展,2007,19:408—412
2. Haruyama T.Micro- and nanobiotechnology for biosensing cellular responses.Adv Drug Deliv Rev, 2003,55:393—401
3. Jain KK.Nanodiagnostics: application of nanotechnology in molecular diagnostics.Expert Rev Mol Diagn,2003,3:153—61
4. Vo-Dinh T,Cullum BM, Stokes DL,Nanosensors and biochips: frontiers in biomolecular diagnostics, Sens Actuators B, 2001,74:2—11
5. Cahn RW.Nanomaterials: synthesis, properties and applications.Nanostructured Materials, 1997,8:377—1379
6. Dowling AP,Development of nanotechnologies.Materials Today,2004,7:30—135
7. Roco MC.Reviews of national research programs in nanoparticle and nanotechnology research: Nanoparticle and nanotechnology research in the U.S.A..Journal of Aerosol Science, 1998,29:749—1760
8.解思深.纳米材料体系物理简介.现代科学仪器,1998(1):5—8
9.刘忆,刘卫华,訾树燕等.纳米材料的特殊性能及其应用.沈阳工业大学学报,2000(1): 2 1—24
10.高春华.纳米材料的基本效应及其应用.江苏理工大学学报(自然科学版), 2001,22:45—49
11.陈月辉,赵光贤.纳米材料的特性和制备方法及应用[J].橡胶工业,2004,51:182—188
12. Pitkethly MJ.Nanomaterials– the driving force.Materials Today.2004,7:20—29
13. Hayat MA.Colloidal Gold: Principles, Methods, and Applications.Academic Press: San Diego, 1991
14. Murray CB.,Norris, D. J..Bawendi.M.G.J.Am.Chem.Soc.,1993,115,8706—8715
15. Feltin N,Pileni MP. Langmuir.1997,13:3927—3933
16. Huang CZ,Li KA,Tong SY.Anal.Chem.,1996,68(13):2259
17. Anglister J,Steinberg IZJ.Chem.Phys.,1981,74(2):786
18. Bock LC, Griffin LC, Latham JA, et al. Selection of single stranded DNA molecules that bind and inhibit human thrombin.Nature,1992,355(6360):564—571
19. Woolley AT.Biomedical microdevices and nanotechnology.Trends in Biotechnology, 200 -1,19:38—39
20. Wei Y, Cao C, Jin RC, et al. Nanoparticles with Raman.Spectroscopic Fingerprints for DN -A and RNA Detection.Science,2002, 297:1536—1540.
21. Li YX,Chen JL,Zhu CQ,et al.Preparation and application of cysteine-capped ZnS nanopar-ticles as fluorescence probe in the determination of nucleic acids.Spectrochimica Acta Part A, 2004, 60:1719—1724
22.Zhang J,Malicka J,Gryczynski I, et al.Oligonucleotide displaced organic monolayer-protec -ted silver nanoparticles and enhanced luminescence of their salted aggregates.Analytical Bio -chemistry, 2004,330: 81—86
23.施利毅.纳米科技基础[M].上海:华东理工大学出版社,2005.133
24.林章碧,苏星光,张家骅等.分析化学,2002,30(2):237—241
25. Hunter RJ.Foundations of Colloid Science.Oxford University Press Inc.: New York, 200 -1.Shaw, D. J. Colloid and Surface Chemistry.Butterworth-Heinemann Ltd.: Oxford, 1991
26. Quinten M,Kreibig US.Science,1986,172: 557
27. Storhoff JJ,Lazarides AA,Mucic RC,et al.Am.Chem.Soc.,2000,122:4640
28.Li HX,Lewis J.Rothberg.Label-Free Colorimetric Detection of Specific Sequences in Genomic DNA Amplified by the Polymerase Chain Reaction.J.AM.Chem.Soc.:2004,126:109 58—10961
29. Maxwell DJ,Taylor JR,Nie SJ.Am.Chem.Soc.2002,124:9606
30. Graham D,Mallinder BJ,Smith WE.Angew.Chem.Int.Ed.,2000,39,1061
31. Bloomfield VA,Crothers DM,Tinoco JRI.Nuclei Acids:Structures,Properties,and Function -s.University Science Books:Sausalito,CA,1999
32. Demers LM, Mirkin CA, Mucic RC.A Fluorescence-Based Method for Determining the Surface Coverage and Hybriaization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles.Anal.Chem.,2000,72: 5535—5541
33. Mirkin CA, Letsinger RL,Mucic PC,et al.Nature,1996,382:607—609
34. Storhoff JJ,Lazarides AA,Mirkin CA,et al.J.Am.Chem.Soc.,2000,122:4640—4650
35. Jin R,Wu G,Li ZJ.Am.Chem.Soc.,2003,125:1643—1654
36. LyttonrJean A KR,Mirkin CA.J.Am.Chem.Soc.,2005,127:12754—12755
37. Demers LM,Ostblom M,Zhang H,et al.J.Am.Chem.Soc.,2002,124:11248—11249
38. Bao P,Frutos AG,Greef C,et al.Anal.Chem.,2002,74:1792—1797
39.Taylor JR.Doctoral Dissertation of Indiana University.USA,2002:122—147
40. Gourishankar A,Shukla S,Ganesh KN,et al.J.Am.Chem.Soc.,2004,126:13186—13187
41. Authier L,Grossiord C,Limoges B,et al.Anal.Chem.,2001,73:4450—4456
42. Park SJ,Taton TA,Mirkin CA.Science,2002,295:1503—1505
43.蔡宏,王延琴,何品刚等.高等学校化学学报,2003,24(8) :1390—1394
44. Zhou XC,O’Sheaa SJ,Li SFY.Chem.Commun.,2000:953—954
45.刘涛,唐季安,韩梅梅等.科学通报,2003,48(4) :342—344
46. Liu T,Tang J,Jiang L.Biochem.Biophys.Res.Commun,2004,313:3—7
47. Hajime S,Michiyo E.Journal of Histochemistry & Cytochemistry,2000,48(4) :493—498
48. Ellington AD,Szostak JW.In-vitro selection of RNA molecules that bind specific liganes. Nature,1990,346(30):818
49. Jiang YX,Zhu CF,Ling LS.Specific aptamer-protein interaction studied by atomic force microscopy.Anal Chem.,2003,75:2112
50. Lee JF,Hesselberth JR,Meyers LA,et al.Aptamer database.Nucleic Acids Res.,2004,32 Database issue:D95
51. Thomas H,D inshaw J,Patel.Adaptive Recognition by Nucleic Acid Aptamers. Science, 2000,287 (5454):820
52. Liss M, Petersen B, Wolf H, et al. An aptamer-based quartz crystal protein biosensor. Anal.Chem.,2002,74:4488—4495
53. Ruckman J, Green LS, Beeson J, et al.2’-fluoropyrimidine RNA-based aptamers to the 165-amino acid form of vascular endothelial growth factor(VEGF165).J.Biol.Chem.,1998, 273(20):556—567
54. Gold LJ.Oligonucleotides as research,diagnostic,and therapeutic agents.Biol.Chem., 199 -5,270:13581
55. Szpechcinski A,Grzanka A.Aptamers in clinical diagnostics.Postepy Biochem,2006,52 (3):260—70
56.王成刚.核酸适配体技术研究进展[J].生物医学工程学杂志,2006,23(2):463—466
57.黄全跃.直接凝血酶抑制剂在急性冠状动脉综合症中的应用[J].中国动脉硬化杂志, 2003,11(2):175—177
58.张莹.凝血酶活性的检测及临床意义[J].微循环学杂志,2005,15(2):70—72
59. Mirkin CA,Letsinger RL,Mucic RC,et al.A DNA-based method for rationally assembling nanoparticles into macroscopic materials.Nature,1996,382(6592) :607—609
60. Elghanian R,Srorhoff JJ,Mucic RC,et al.Selective colorimetric detection of polynuclcotid -es based on the distance-dependent optical properties of gold nanoparticles.Science,1997,27 -7(5329):1078—1081
61. ReynoldsⅢRA, Mirkin CA,Letsinger RL.A gold nanoparticle/latex microsphere-based colorimetric oligonuclcotide detection method.Pure Appl Chem.,2000,72(1-2):229—335
62. Taton TA,Mirkin CA,Letsinger RL.Scanometric DNA array detection with nanoparticle probe. Anal Chem.,2000,289(5485):1757—1760
63. Nie L,Tang J,Guo H,et al.Colorimetric detection of polynuclcotides on polypropylene -slices.Anal Sci,2004,20(3):461—463
64. Cao YC,Jin R,Mirkin CA.Nanoparticles with Ramam spectroscopic fingerprints for DNA and RNA detection.Science,2002,297(5586):1536—1540
65. Sato K,Hosokawa K,Macda M.Rapid aggregation of gold nanoparticles induced by non-c -ross-linking DNA hybridization.J Am Chem Soc,2003,125(27):8102—8103
66.乐晓萍,杜鹏,张钦宪等.聚丙烯酰胺凝胶银染技术改良[J].河南医科大学学报,2001,36 (4):395
67.王美佳,纪小会,王连英等.DNA在纳米金标上的组装、杂交、检测与银增强[J].物理化学学报.2003,19(9):879—882
68.赵立凡,李柏生,黑笑涵等.银染增强的纳米金探针技术检测微量核酸[J].中国生物化学与分子生物学报.2006,11:919—923
69. Pavlov V, Xiao Y,Shlyahovsky B,et al.Aptamer-Functionalized Au Nanoparticles for the Amplified Optical Detection of Thrombin.J Am Chem Soc.,2004,126:11768—11769
70.韩鹤友,崔华,林祥钦.化学发光分析法应用新进展[J].光谱实验室.2002,19(1):39—40
71.方肇伦等.流动注射分析法[M].北京:科学出版社,1999,1—4
72. Ruzicka J, Hansen EH.Flow Injection Analysis(-ndEd).New York: John Wiley&Sons,198 -8
73.李原芳,黄承志,胡小莉.分析化学,1998,26(12):1508
74. Stanton SG,Decora RJJ.Chem.Phys.,1981,75:5615
75. Su X, Chew FT, Li SFY.Design and application of piezoelectric quartz crystal-based immunoassay.Anal.Sci.,2000,16:107—14
76. Liu T, Tang J, Jiang L.The enhancement effect of gold nanoparticles as a surface modifier on DNA sensor sensitivity.Biochem Biophys Res Commun,2004,313:3—7
77. Gulietti A,Overbergh L,Valckx D,et al.An overview of real-time quantitative PCR:applic -ations to quantify cytokine gene expression.Methods,2001,25(4):386—401
78. Tasset DM,Kublik MF, Steiner W,et al.Oligonucleotide inhibitor of human thrombin that bind distinct epitopes.Journal of Molecular Biology,1997,272(5):688—698
79.汪治清,佟玉品,杨新科.Aptamer核酸药物的研究进展[J].中华实验和临床病毒学杂志,2002,16(3):295—298
80. Spiridonova VA,RogE V,Dugina TN,et al.Aptamer DNA-a new type of thrombin inhibito -r.Bioorg Khim,2003,29(5):495—498
81.蔡强,吴维明,陈裕泉.纳米金粒子的生物组装技术与光学检测技术研究[J].传感技术学报.2003.3(1):22
82. Li ZP,Duan XR,Liu CH, et al.Selective determination of cysteine by resonance light scatte -ring technique based on self-assembly of gold nanoparticles.Analytical Biochemistry,2006,35 -1:18—25
83.Du BA,Li ZP,Liu CH.One-step homogeneous detection of DNA hybridization with gold nanoparticle probe by using a linear light-scattering techniqu e.Angew.Chem.Int.Ed.,2006,4 -5:8022—8025
2. Haruyama T.Micro- and nanobiotechnology for biosensing cellular responses.Adv Drug Deliv Rev, 2003,55:393—401
3. Jain KK.Nanodiagnostics: application of nanotechnology in molecular diagnostics.Expert Rev Mol Diagn,2003,3:153—61
4. Vo-Dinh T,Cullum BM, Stokes DL,Nanosensors and biochips: frontiers in biomolecular diagnostics, Sens Actuators B, 2001,74:2—11
5. Cahn RW.Nanomaterials: synthesis, properties and applications.Nanostructured Materials, 1997,8:377—1379
6. Dowling AP,Development of nanotechnologies.Materials Today,2004,7:30—135
7. Roco MC.Reviews of national research programs in nanoparticle and nanotechnology research: Nanoparticle and nanotechnology research in the U.S.A..Journal of Aerosol Science, 1998,29:749—1760
8.解思深.纳米材料体系物理简介.现代科学仪器,1998(1):5—8
9.刘忆,刘卫华,訾树燕等.纳米材料的特殊性能及其应用.沈阳工业大学学报,2000(1): 2 1—24
10.高春华.纳米材料的基本效应及其应用.江苏理工大学学报(自然科学版), 2001,22:45—49
11.陈月辉,赵光贤.纳米材料的特性和制备方法及应用[J].橡胶工业,2004,51:182—188
12. Pitkethly MJ.Nanomaterials– the driving force.Materials Today.2004,7:20—29
13. Hayat MA.Colloidal Gold: Principles, Methods, and Applications.Academic Press: San Diego, 1991
14. Murray CB.,Norris, D. J..Bawendi.M.G.J.Am.Chem.Soc.,1993,115,8706—8715
15. Feltin N,Pileni MP. Langmuir.1997,13:3927—3933
16. Huang CZ,Li KA,Tong SY.Anal.Chem.,1996,68(13):2259
17. Anglister J,Steinberg IZJ.Chem.Phys.,1981,74(2):786
18. Bock LC, Griffin LC, Latham JA, et al. Selection of single stranded DNA molecules that bind and inhibit human thrombin.Nature,1992,355(6360):564—571
19. Woolley AT.Biomedical microdevices and nanotechnology.Trends in Biotechnology, 200 -1,19:38—39
20. Wei Y, Cao C, Jin RC, et al. Nanoparticles with Raman.Spectroscopic Fingerprints for DN -A and RNA Detection.Science,2002, 297:1536—1540.
21. Li YX,Chen JL,Zhu CQ,et al.Preparation and application of cysteine-capped ZnS nanopar-ticles as fluorescence probe in the determination of nucleic acids.Spectrochimica Acta Part A, 2004, 60:1719—1724
22.Zhang J,Malicka J,Gryczynski I, et al.Oligonucleotide displaced organic monolayer-protec -ted silver nanoparticles and enhanced luminescence of their salted aggregates.Analytical Bio -chemistry, 2004,330: 81—86
23.施利毅.纳米科技基础[M].上海:华东理工大学出版社,2005.133
24.林章碧,苏星光,张家骅等.分析化学,2002,30(2):237—241
25. Hunter RJ.Foundations of Colloid Science.Oxford University Press Inc.: New York, 200 -1.Shaw, D. J. Colloid and Surface Chemistry.Butterworth-Heinemann Ltd.: Oxford, 1991
26. Quinten M,Kreibig US.Science,1986,172: 557
27. Storhoff JJ,Lazarides AA,Mucic RC,et al.Am.Chem.Soc.,2000,122:4640
28.Li HX,Lewis J.Rothberg.Label-Free Colorimetric Detection of Specific Sequences in Genomic DNA Amplified by the Polymerase Chain Reaction.J.AM.Chem.Soc.:2004,126:109 58—10961
29. Maxwell DJ,Taylor JR,Nie SJ.Am.Chem.Soc.2002,124:9606
30. Graham D,Mallinder BJ,Smith WE.Angew.Chem.Int.Ed.,2000,39,1061
31. Bloomfield VA,Crothers DM,Tinoco JRI.Nuclei Acids:Structures,Properties,and Function -s.University Science Books:Sausalito,CA,1999
32. Demers LM, Mirkin CA, Mucic RC.A Fluorescence-Based Method for Determining the Surface Coverage and Hybriaization Efficiency of Thiol-Capped Oligonucleotides Bound to Gold Thin Films and Nanoparticles.Anal.Chem.,2000,72: 5535—5541
33. Mirkin CA, Letsinger RL,Mucic PC,et al.Nature,1996,382:607—609
34. Storhoff JJ,Lazarides AA,Mirkin CA,et al.J.Am.Chem.Soc.,2000,122:4640—4650
35. Jin R,Wu G,Li ZJ.Am.Chem.Soc.,2003,125:1643—1654
36. LyttonrJean A KR,Mirkin CA.J.Am.Chem.Soc.,2005,127:12754—12755
37. Demers LM,Ostblom M,Zhang H,et al.J.Am.Chem.Soc.,2002,124:11248—11249
38. Bao P,Frutos AG,Greef C,et al.Anal.Chem.,2002,74:1792—1797
39.Taylor JR.Doctoral Dissertation of Indiana University.USA,2002:122—147
40. Gourishankar A,Shukla S,Ganesh KN,et al.J.Am.Chem.Soc.,2004,126:13186—13187
41. Authier L,Grossiord C,Limoges B,et al.Anal.Chem.,2001,73:4450—4456
42. Park SJ,Taton TA,Mirkin CA.Science,2002,295:1503—1505
43.蔡宏,王延琴,何品刚等.高等学校化学学报,2003,24(8) :1390—1394
44. Zhou XC,O’Sheaa SJ,Li SFY.Chem.Commun.,2000:953—954
45.刘涛,唐季安,韩梅梅等.科学通报,2003,48(4) :342—344
46. Liu T,Tang J,Jiang L.Biochem.Biophys.Res.Commun,2004,313:3—7
47. Hajime S,Michiyo E.Journal of Histochemistry & Cytochemistry,2000,48(4) :493—498
48. Ellington AD,Szostak JW.In-vitro selection of RNA molecules that bind specific liganes. Nature,1990,346(30):818
49. Jiang YX,Zhu CF,Ling LS.Specific aptamer-protein interaction studied by atomic force microscopy.Anal Chem.,2003,75:2112
50. Lee JF,Hesselberth JR,Meyers LA,et al.Aptamer database.Nucleic Acids Res.,2004,32 Database issue:D95
51. Thomas H,D inshaw J,Patel.Adaptive Recognition by Nucleic Acid Aptamers. Science, 2000,287 (5454):820
52. Liss M, Petersen B, Wolf H, et al. An aptamer-based quartz crystal protein biosensor. Anal.Chem.,2002,74:4488—4495
53. Ruckman J, Green LS, Beeson J, et al.2’-fluoropyrimidine RNA-based aptamers to the 165-amino acid form of vascular endothelial growth factor(VEGF165).J.Biol.Chem.,1998, 273(20):556—567
54. Gold LJ.Oligonucleotides as research,diagnostic,and therapeutic agents.Biol.Chem., 199 -5,270:13581
55. Szpechcinski A,Grzanka A.Aptamers in clinical diagnostics.Postepy Biochem,2006,52 (3):260—70
56.王成刚.核酸适配体技术研究进展[J].生物医学工程学杂志,2006,23(2):463—466
57.黄全跃.直接凝血酶抑制剂在急性冠状动脉综合症中的应用[J].中国动脉硬化杂志, 2003,11(2):175—177
58.张莹.凝血酶活性的检测及临床意义[J].微循环学杂志,2005,15(2):70—72
59. Mirkin CA,Letsinger RL,Mucic RC,et al.A DNA-based method for rationally assembling nanoparticles into macroscopic materials.Nature,1996,382(6592) :607—609
60. Elghanian R,Srorhoff JJ,Mucic RC,et al.Selective colorimetric detection of polynuclcotid -es based on the distance-dependent optical properties of gold nanoparticles.Science,1997,27 -7(5329):1078—1081
61. ReynoldsⅢRA, Mirkin CA,Letsinger RL.A gold nanoparticle/latex microsphere-based colorimetric oligonuclcotide detection method.Pure Appl Chem.,2000,72(1-2):229—335
62. Taton TA,Mirkin CA,Letsinger RL.Scanometric DNA array detection with nanoparticle probe. Anal Chem.,2000,289(5485):1757—1760
63. Nie L,Tang J,Guo H,et al.Colorimetric detection of polynuclcotides on polypropylene -slices.Anal Sci,2004,20(3):461—463
64. Cao YC,Jin R,Mirkin CA.Nanoparticles with Ramam spectroscopic fingerprints for DNA and RNA detection.Science,2002,297(5586):1536—1540
65. Sato K,Hosokawa K,Macda M.Rapid aggregation of gold nanoparticles induced by non-c -ross-linking DNA hybridization.J Am Chem Soc,2003,125(27):8102—8103
66.乐晓萍,杜鹏,张钦宪等.聚丙烯酰胺凝胶银染技术改良[J].河南医科大学学报,2001,36 (4):395
67.王美佳,纪小会,王连英等.DNA在纳米金标上的组装、杂交、检测与银增强[J].物理化学学报.2003,19(9):879—882
68.赵立凡,李柏生,黑笑涵等.银染增强的纳米金探针技术检测微量核酸[J].中国生物化学与分子生物学报.2006,11:919—923
69. Pavlov V, Xiao Y,Shlyahovsky B,et al.Aptamer-Functionalized Au Nanoparticles for the Amplified Optical Detection of Thrombin.J Am Chem Soc.,2004,126:11768—11769
70.韩鹤友,崔华,林祥钦.化学发光分析法应用新进展[J].光谱实验室.2002,19(1):39—40
71.方肇伦等.流动注射分析法[M].北京:科学出版社,1999,1—4
72. Ruzicka J, Hansen EH.Flow Injection Analysis(-ndEd).New York: John Wiley&Sons,198 -8
73.李原芳,黄承志,胡小莉.分析化学,1998,26(12):1508
74. Stanton SG,Decora RJJ.Chem.Phys.,1981,75:5615
75. Su X, Chew FT, Li SFY.Design and application of piezoelectric quartz crystal-based immunoassay.Anal.Sci.,2000,16:107—14
76. Liu T, Tang J, Jiang L.The enhancement effect of gold nanoparticles as a surface modifier on DNA sensor sensitivity.Biochem Biophys Res Commun,2004,313:3—7
77. Gulietti A,Overbergh L,Valckx D,et al.An overview of real-time quantitative PCR:applic -ations to quantify cytokine gene expression.Methods,2001,25(4):386—401
78. Tasset DM,Kublik MF, Steiner W,et al.Oligonucleotide inhibitor of human thrombin that bind distinct epitopes.Journal of Molecular Biology,1997,272(5):688—698
79.汪治清,佟玉品,杨新科.Aptamer核酸药物的研究进展[J].中华实验和临床病毒学杂志,2002,16(3):295—298
80. Spiridonova VA,RogE V,Dugina TN,et al.Aptamer DNA-a new type of thrombin inhibito -r.Bioorg Khim,2003,29(5):495—498
81.蔡强,吴维明,陈裕泉.纳米金粒子的生物组装技术与光学检测技术研究[J].传感技术学报.2003.3(1):22
82. Li ZP,Duan XR,Liu CH, et al.Selective determination of cysteine by resonance light scatte -ring technique based on self-assembly of gold nanoparticles.Analytical Biochemistry,2006,35 -1:18—25
83.Du BA,Li ZP,Liu CH.One-step homogeneous detection of DNA hybridization with gold nanoparticle probe by using a linear light-scattering techniqu e.Angew.Chem.Int.Ed.,2006,4 -5:8022—8025
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |