摘要
脱氧核糖核酸(DNA)被称为生命体内的遗传物质,研究发现人类的许多遗传疾病都与DNA分子中碱基序列的变异有关。检测与疾病有关的变异对基因筛选、遗传疾病的早期诊断和治疗具有十分深远的意义。
DNA杂交分析技术是目前生物化学和分子生物学研究中应用最广泛的技术之一,是定性或定量检测特异DNA序列片段的有力工具。目前,它已广泛应用在生命科学,尤其是医学的各个领域。传统的DNA分子杂交采用的是放射性标记的检测方法,这种方法虽然灵敏度高,但存在放射性物质对人体及环境的危害。自20世纪80年代以来,各种非同位素如酶、荧光素、生物素、地高辛标记的化学发光法和荧光分析法以及以电活性物质做标记的电化学方法相继问世,这些方法虽然在一定程度上克服了同位素标记的缺陷,但由于存在灵敏度不够高或检测系统庞杂或仪器价格昂贵或标记物不稳定等缺陷,还不能完全取代传统方法。因此寻求简单、灵敏、可靠、价廉的非放射性标记的DNA检测方法具有十分重要的现实意义。
电致化学发光(ECL)分析是一种通过电极反应而产生化学发光的微量分析方法。它将电化学技术与化学发光检测结合起来,弥补了电化学方法和化学发光方法的不足,具有灵敏度高、选择性好、动力学范围宽、检测快速方便、仪器价格便宜等优点,近几年来在分析化学,尤其在生物分析领域引起了人们的极大关注。本论文通过研究了多种ECL活性物质的发光性能,并以这些物质为标记物制备了多种高灵敏度的DNA-ECL探针,结合DNA杂交技术和DNA固定化技术,将高灵敏度的ECL检测手段应用于生命物质DNA的序列识别及含量测定,为DNA传感器的研究和基因芯片的开发提供了新的思路和方法。
摘要
论文主要研究内容如下:
第一章绪论
简单介绍了ECL分析方法的概念、原理和特点,重点介绍了各种ECL反应
类型及其在分析化学中的应用,系统总结了各种ECL传感器,展望了ECL分析
的未来发展方向。介绍了DNA传感器的构造和各种DNA传感器的原理、特点
及分析应用。最后阐述了本论文的目的和意义,指出论文的创新之处及主要研究
内容。
第二章ECL活性标记物的性能研究及其ECL分析体系的建立
研究了环酞麟类物质N一(4一氨基丁基)一乙基异鲁米诺(ABEI)和鲁米诺
(Luminol)、叮淀酷类物质醛基叮淀酷和竣基叮睫酷、抗癌药物道诺霉素(DNR)
的ECL行为,建立了这几种物质的ECL分析体系,并结合电化学、紫外一可见
光谱、荧光光谱等手段讨论了它们的ECL反应机理,为DNA的ECL分析奠定
了方法基础。
1 ABEI的ECL性能研究及其ECL分析体系的建立
ABEI是一种环酞麟类衍生物,其分子结构上含有一个特殊的长臂功能团一氨
基,可直接用来标记生物大分子。本文研究了ABEI的ECL性能,发现在碱性
溶液中,施加+l .ov(vsA留AgCI)的电压时,ABEI可在Pt电极上产生很强的
ECL信号,适量HZO:的加入可催化这一发光反应。考察了各种影响因素,建立
了ABEI的ECL分析方法.方法测定ABEI的线性响应范围为1 .3 x 10石一6.5x
10·’Zmol/L,检出限可以达到9.6xz0·’3mo比。
2 FCA催化Luminol一HZO:ECL反应的研究及其ECL分析体系的建立
Luminol是最早被发现的ECL物质,也是目前为止发光效率最高的物质之一,
但由于其分子结构上缺乏合适的标识基团而使其难以直接标记生物大分子。梭基
二茂铁(FCA)是一种电化学活性物质,其分子结构上的梭基能够和生物大分子
摘要
作用,可用于标记生物大分子。本文研究了FCA对Luminol一HZOZ的ECL催化作用,
发现在低电位下,FCA可极大地催化Luminol和HZOZ的ECL反应,而且催化信号
与FCA的浓度在5.0、10一9一2.0x10礴moFL的范围内呈良好的线性关系,检出限可
以达到1.5、10一gmol几。以FCA作为DNA标记物,方法可用于DNA的ECL检测。
3叮睫酷类化合物的ECL性能研究及其ECL分析体系的建立
叮咙醋是一类较好的化学发光(CL)物质,而且其分子结构上含有特殊的
功能团,可用来标记生物大分子,实现生物物质的CL分析。但有关其ECL性
能的研究却较少。本文研究了碘化10一乙梭基叮咙一9一梭酸苯醋(C AcPEI)和对甲
酞基苯基一10一甲基叮咤一9一梭酸酷氟磺酸盐(MFPA)的ECL行为,发现在中性
KNO3介质中,这两种叮咙醋都能在一定的电位下产生很强的ECL信号,考察
了各种影响因素,确定了发光的最佳条件,探讨了可能的发光机理。在最佳条件
下EeL信号与eAePEx及MF琳的浓度分别在5.oxlo.,o一l.Zxlo一79/ml和
2.4xl0‘’0一9.6xlo一8留ml范围内呈良好的线性关系。检出限可分别达到2.4只10一’“
g加l和l.6xlo一’“g/ml。比eL法的灵敏度高1000倍。
4道诺霉素的
道诺霉素
ECL性能研究及其ECL分析体系的建立
(DNR)是一种葱环类抗生素,
上得到广泛的应用
可选择性地嵌入到
。与DNA作用机理研究表明
具有广谱的抗肿瘤作用,在临床
,DNR以其特殊的葱环平面结构
DNA
研究了DNR的ECL行为
的双链结构区。因此可作为DNA的杂交指示剂。本文
,发现在
Deoxyribonucleic Acid (DNA) is known as inherited substance in life body. With the increasing of knowledge about human beings diseases it is found that the mutations of base sequence in DNA are responsible for numerous inherited human disorders. In order to make early diagnosis and cure for disease, much effort is needed to detect the specific DNA sequences related to diseases, which is also helpful for the selection of genes and the development of drugs.
Most of the conventional methods for the analysis of specific DNA sequences are based on the nucleic acid hybridization. Single-stranded ss- DNA probe binds to its complementary strand (target sequence) to form a double-stranded (ds) DNA with high efficiency. The signal provided by the probe is a measure of the degree of hybridization.
Traditional DNA probes are labeled with radioisotopes i.e. 32P, I25I, 3H or 35S. Although this method gives high sensitivity, the radioactive labels present many problems such as a potential hazard to analyst and environment, which limited its application in DNA diagnostic laboratories. In order to overcome these problems a serious of non-radioactive DNA probes such as fluorescent, chemiluminescent and electrochemical probes have been developed. Although these new methods display many advantages, they have not been used to take place completely the traditional method because of low sensitivity or complex equipment or other shortcomings. So it is necessary to develop other rapid and sensitive methods for the detection of specific DNA sequences, which can also be used easily in non-specialized laboratories. Electrochemiluminescence (ECL) is a technique that combines chemiluminescence
(CL) and electrochemistry (EC). The CL reaction was initiated by an electrochemical reaction at electrode surface. Such an electro-initiation reaction introduces a large number of additional advantages, such as high sensitivity and selectivity, rapid and convenient operation and relatively simple instrumentation system. Due to these inherent advantages, ECL method has attracted much attention from all analytical fields, especially from biochemical analysis. In this dissertation we focused on the preparation of a new type of DNA probes which were labeled with ECL activated substances. Based on coupling with the DNA hybridization and immobilization techniques, we have developed new ECL methods for the determination of special DNA sequence. Compared with other methods the new methods are simple, rapid as well as sensitive and selective. The dissertation is composed of four chapters as folio wings:
Chapter 1: Introduced the ECL and its application in analytical chemistry field, paid more attention to the types of ECL reaction and ECL sensors. Reviewed the DNA sensors, including their principles, advantage and disadvantage. At last pointed out the purpose of the dissertation.
Chapter 2: Study on the ECL behaviors of several substances and establishment of ECL analytical systems
1 The ECL behavior of N-(4-aminobutyl)-N-ethylisoluminol (ABEI) was studied and found ABEI could produce emission light when oxidized at a +1.0 V (vs Ag/AgCl) potential in alkaline solution. The addition of H_(2)O_(2) markedly improved the ECL sensitivity. The effected factors on the ECL were examined. Under optimal conditions, ABEI took a linear response to ECL intensity in the range of 1.3x10-6 ~ 6.5 xl0-12 mol/L. The detection limit is 2.2 xl0-12 mol/L at a signal to noise ratio of 3.
2 The catalytic reaction of ferrocenecarboxylic acid (FCA) to Luminol and hydrogen peroxide was investigated. It was found the Chemiluminescent intensity of
Luminol with hydrogen peroxide can be greatly enhanced by electrochemically oxidizing of FCA at low potential in Tris buffer solution. Under optimal conditions the ECL intensity exhibits a good linear response with the concentration of FCA in the range of 5.0x10~(-9).~2.0xl0-4 mol/L. The detection limit is 1.5xl0-9mol/L.
3 A study on the ECL of acridinum esters
引文
1 Harvey N., J. Phys. Chem., 1929, 33: 1456
2 Kuwana T., Epstein B., Seo E., J. Phys. Chem., 1963,67: 2243
3 Kuwana T.,J. Electroanal. Chem., 1963, 6: 164
4 Schaper H., J. Electroanal. Chem. Interfacial Electrochem., 1981,129: 335
5 Blatchford C., and Malcolme-Lawes D. J., J. Chromatogr., 1985,227: 321
6 Blatchford C., Humphreys E., and Malcolme-Lawes D. J., J. Chromatogr., 1985, 281:329
7 Nieman T. A., Mikrochem. Acta, 1980, 3: 239
8 Nieman T. A., J. Res. Natl. Bur. Stand., 1988, 93: 501
9 Wang Y., and Yeung E. S., Anal Chim. Acta, 1992, 295:266
10 Downcy T. M., and Nieman T. A., Anal. Chem., 1992, 64:261
11 Litting J. S. and Nieman T. A., Anal. Chem., 1992, 64:1140
12 Laespada M. E. F., Pavon J. L. P., Cordero B. M., Anal. Chim. Acta, 1996, 327:253
13 Marquette C. A., Blum L. J., Sens. Actuators B, 1998, 51:100
14 Steijger O. M., Kmminga D. A., Brummelhuis, Lingeman H., J. Chromatogr. A,1998, 799:57
15 王鹏,张文艳,周泓,朱果逸《分析化学》1998,26(7):898
16 安镜如,林建国《化学学报》1992,50,594
17 庄惠生,王琼娥,张帆《分析化学》1997,25(3),330
18 Yasuo Y., Hirotak H., Takehior M., J. Chem. Engineer., Jap., 1996, 29(5): 851
19 Aizawa M., Pro. BCEIA. Electroanal. Chem., 1997, F25
20 Blackburn G. F., Shah H. P., Kenten J. H., Clin. Chem., 1991, 37(9): 1534
21 Daniel R. D., Nature, 1995, 377:758
22 Nicbolas R., Hoyle N. R., Eckert B., Clin. Chem., 1996, 42(9): 1576
23 Wellerl L., Kenten., E., Freeman, Clin. Chem., 1995, 41(6): S221
24 Bialk P., Vogel. R., Mayr S., Clin. Chem., 1995, 41(6): S60
25 Bard A. J., Whiteside G M., US Patent, 1994, 5310687
26 Kankare J., Haapakka K., Kulmala S. et al., Anal. Chim. Acta, 1992, 266:205
27 Michel P. E., de Rooij N. F., Koudelka-Hep M., Fahnrich K. A., O'Sullivan C. K.,Guilbault G.G., J. Electroanal. Chem., 1999, 474:192
28 Kim J. H., Shin H. B., Park K. I., Lee H. M., Kwon O. H., Kim D. S., Clin. Chem.,1998, 44:A124
29 王鹏,张文艳,周泓,朱果逸《科学通报》1998,43(21):2241
30 周永列,吕亚萍《国外医学临床生物化学与检验学分册》1998,19(4):149
31 Kenten J. H., Casade J., Link. J., et al., Clin. Chem., 1991, 37:1626
32 Dicesare J., Grossman B., Karz E., et al., Biotechniques, 1993, 15:152
33 Hsueh Y. T., Collins S. D., Smith R. L., Sens. and Actuators, 1998, B49:1
34 Hsueh Y. T., Northrup M. A., Smith R. L., Sens. and Actuators, 1996, B33:110
35 Motmans K., Raus J., Vandevyver C., J. Immunol. Methods, 1996, 190:107
36 O'Connell C. D., Juhasz A., Kuo C., Reeder D. J., Hoon D. S. B., Clin. Chem., 1998, 44:1161
37 安镜如,林金明,陈曦《分析化学》1991,19(11):1340
38 陈曦,王小如,黄本立《分析化学》1998,26(6):770
39 王鹏,袁艺,朱国逸,张密林《分析化学》1999,27(10):1219
40 Greenway G M., Trends Anal. Chem., 1990, 9:200
41 Knight A. W., Greenway G. M.,Analyst, 1994, 119:879
42 Knight A. W., Trends Anal. Chem., 1999,18:47
43 Fahnrich K. A., Pravda M., Guilbault G G, Tanlanta, 2001, 54:531
44 Marcus R. A., J. Chem. Phys., 1965, 43:2654
45 Feldburg S. W., J. Am. Chem. Soc., 1966, 88:390
46 Kim J., Faulker L. R., J. Electroanal. Chem. Interfacial Electrochem., 1988, 242: 107
47 Bard A. J., Faulker L. R., Electrochemical Methods, Fundamentals and Applications, New York: Wiley, 1980:621
48 Happakka K., Kankare J., and Lipiainen K., Anal. Chim. Acta, 1988, 215:341
49 Richard T. C., Bard A. J., Anal. Chem., 1995, 67:3140
50 Collinson M. M., Wightman R. M., Anal. Chem., 1993, 65:2579
51 林祥钦,孙玉刚,崔华《分析化学》1999,27(5):497
52 安镜如,陈曦《高等学校化学学报》1989,10(11):1110
53 Haapakka K. E., and Kankare J. J.,Alal. Chim. Acta, 1982, 138:253
54 Haapakka K. E., and Kankare J. J.,Anal. Chim. Acta, 1982, 138:263
55 Vitt J. E., Johnson D. C., and Engstrom R. C., J. Electrochem. Soc., 1991, 138: 1637
56 Nieman T. A., J. Res. Stand., Sect. A, 1988, 93:501
57 Sakura S., Anal. Chim. Acta., 1992, 262:49
58 Jirka G. P., Martin A. F., Nieman T. A., Anal. Chim. Acta, 1993, 284:345
59 Keamey N. J., Hall C. E., Jewsbury R. A., Timmis S. G, Anal. Commun., 1996, 7:815
60 Leca B., Blum L. J., Analyst, 2000, 125: 789
61 Sakura S., Imai H., Anal. Sci., 1988, 4: 9
62 Marquette C. A., Blum L. J., Anal. Chim. Acta, 1999, 381: 1
63 Atwater J. E., Akse J. R., DeHart J., Wheeler R. R., Anal. Lett., 1997, 30 (1) : 1
64 Atwater J. E., Akse J. R., DeHart J., Wheeler R. R., Anal. Lett., 1997, 30 (8) : 1445
65李瑛秀, 朱连德, 朱果逸《分析试验室》2001,20(5) :89
66 Zhu L. D., Li Y. X., Zhu G Y, Sens, and Actuators , 2002, B86: 209
67 Tsafack V. C., Marquette C. A., Leca B., Blum L. J., Analyst, 2000,125: 151
68 Marquette C. A., Ravaud. S., Blum L. J., Anal. Lett., 2000, 33 (9) : 1779
69 Hostis E. L., Michel P. E., Fiaccabrino G C., Strike D. J., de Rooij N. F., Koudelka-Hep M., Sens. Actuators, 2000, 64B: 156
70 Kremeskoter J., Wilson R., Schiffrin D. J., Luff B. J., and Wilkinson, Meas. Sci. Technol.,1995,6: 1325
71 Haapakka K. E., and Kankare J. J., Anal. Chim. Acta, 1980, 118: 333
72 Haapakka K. E.,Anal. Chim. Acta, 1982,139: 229
73 Haapakka K. E.,Anal. Chim. Acta, 1982, 141: 263
74安镜如, 姚晴《分析化学》1990,18(9) :867
75 Lasovsky J., and Grambal F., Coll. Czech. Chem. Commun., 1983,48: 477
76 Zhang C., Zhang S., Zhang Z., Analyst, 1998, 123: 1383
77陈扬, 陆祖宏《化学通报》2001,(9) :537
78 Kricka L J., Clin. Chem., 1991, 37(9) : 1471
79 Kawasaki T., Maeda M., and Tsuji A., J. Chromatorgra., 1985,328: 121
80 Yang M. L, Liu C. Z, Qian K. J, He P. G, Fang Y. Z, Analyst, 2002, 127: 1267
81 Lee W. Y., Mikrochim. Acta, 1997,127: 19
82 Gerardi R. G, Barnett N. W, Lewis S. W., Anal. Chim. Acta, 1999, 378: 1
83 Abruna H. D., J. Electrochem. Soc., 1985, 132: 842
84 Lee C., Ouyang J., Bard A. J., Electroanal. Chem. Interfacial Electrochem., 1988, 244:319
85 Xu G, Dong S., Anal. Chim. Acta, 2000, 12; 235
86 Kane-Maguire N. A. P., Gucker J. A., O'Neil P. J., Inorg. Chem., 1987, 26: 2340
87 Taverna P. J., Mayfield H., Andrews A. R. J., Anal. Chem. Acta, 1998, 373: 111
88 Tokel-Takvoryan N. E., Bard A. J., Chem. Phys. Lett., 1974,25(2) : 235
89 Bonafede S., Ciano M., Bolletta R, Balzani V., Chassot L., Zelewsky A, J. Phys. Chem., 1986,90:3836
90 Richter M. M., Debad J. D., StriplinD. R., Crosby G A., Bard. A. J., Anal. Chem., 1996,68: 4370
91 Vogler A, Kunkely H., Angew. Chem. Int. Ed Engl., 1981,20: 469
92 Vogler A, Kunkely H.,Am. Chem. Soc. Symp. Ser., 1987, 333: 155
93 Mussel R. D., Nocera D. G, J. Am. Chem. Soc., 1988,110: 2764
94 Mussel R. D., Nocera D. G, J. Phys. Chem., 1991, 95: 6919
95 Hemingway R. E., Park S. M., Bard A. J., J. Am. Chem. Soc., 1975, 97: 200
96 Hercules D. M., Lytle F. E., J. Am. Chem. Soc., 1966, 88: 4745
97 Tachikawa H., Bard A. J.,J. Chem. Phys. Lett., 1973,19: 287
98 Tokel N. E., Bard A. J., J. Am. Chem. Soc., 1972, 94; 2862
99 White H. S., Bard A. J., J. Am. Chem. Soc., 1982,104; 6891
100 Rubinstein L, Bard A. J.,J.Am. Chem. Soc., 1981,103: 512
101 Ege D., Becker W. G, Bard A. J., J. Am. Chem. Soc., 1984, 56: 2413
102 Rubinstein L, Martin C. R., Bard A. J., Anal. Chem., 1983, 55: 1580
103 Zu Y., Bard A. J., Anal. Chem., 2000, 72: 3223
104 Leland J. K., and Powell M. J., J. electrochem. Soc., 1990,137 (10) : 3127
105 Noffsinger J. B., and Danielson N. D., Anal. Chem. 1987, 59: 865
106 He L., Cox K. A., Danielson N. D., Anal. Lett., 1990,23:195
107 Brune S. N., and Bobbitt D. R.,Anal. Chem., 1992, 64: 166
108 Chen X., Sato M., J. Flow Inf. Anal., 1995,12:216
109陈曦,佐藤昌宪《分析科学学报》1998,14(4) :278
110陈曦,李金梅,李真,易长青,王小如《分析化学》2002,30(5) :513
111陈曦,易长青,李金梅,李真,王小如《化学学报》2002,60(9) :1662
112 Noffsinger J. B., and Danielson N. D., J. Chromatogr., 1987, 387: 520
113 Arora A., de Mello A. J., Manz. A., Anal. Commun., 1997, 34: 393
114 Clifdon-Hadley F. A., Rev. Med. Microbiol., 2000,11: 47
115 Yu H., Raymonda J. W., McMahon T. M., Campagnari A. A., Biosens. Bioelectron., 2000, 14: 829
116邢婉丽,晁福寰,蒋中华,马立人《高等学校化学学报》,2000,21(6) : 873
117 Grimshaw C., Gleason C., Chojnicki E., Yong J., J. Pharmaceut. Biomed. Anal., 1997, 16: 605
118 Xu X. H., Yang H. C., Mallouk T. E., Bard. A. J., J. Am. Chem. Soc., 1994, 116: 8386
119 Xu X. H., Bard. A. J., J. Am. Chem. Soc., 1995, 117: 2627
120 Stern H. J., Carlos R.D., Schutzbank T. E., Clin. Biochem., 1995, 28: 470
121 Jameison F., Sanchez R. I., Dong L., Leland J. K., Yost D., and Martin M. T., Anal. Chem., 1996, 68: 1298
122 Martin A. F., and Nieman T. A., Biosens. Bioelectro., 1997, 12(6) : 479
123 Chen X., Jia L., Wang X. R., Hu G. L., Sato M., Anal Sci., 1997, 13: 71
124 Chen X., Sato M., Lin Y. J., Microchem. J, 1998, 58; 13
125陈曦,贾丽,佐藤昌宪,《化学学报》,1998,56(3) :238
126 Chen X., Sato M., Anal. Sci. 1995,11: 749
127 Kulmala S., Alk-Kulmala T., Vare L., Helin M., Lehtinen T., Anal. Chim. Acta, 1999, 398: 41
128 Fabrizio E. F., Prieto I., Bard A. J., J. Am. Chem. Soc., 2000, 122: 4996
129 Kulmala S., Kulmala A., Alk-Kleme T., Pihlaja J., Anal. Chim. Acta, 1998, 367: 17
130 Gaillard F., Sung Y. E., Bard A. J.,J. Phys. Chem. B, 1999, 103: 667
131 Kulmala S., Alk-Kulmala T., Kulmala A., Papkovsky D., Loikis K., Anal. Chem, 1998,70: 1112
132 Ala-Kleme T., Latva M., Haapkaak K., Anal Chim Acta, 2000,403: 161
133 Ala-Kleme T., Haapkaak K., Latva M.,J. Alloy. Compd., 1998, 275-277: 911
134 Ala-Kleme T., Klumala S., Latva M., Acta. Chem. Scand., 1997, 51; 541
135 Klumala S., Ala-Kleme T., Joela H., Kulama A., J. Radioanal. Nucl. Chem., 1998, 232: 91
136 Kankare J., Haapakka K., Kulmala S., N(?)nt(?) V., Eskola J., Takola H., Anal. Chim.Acta, 1992, 266:205
137 Kaapkaak K., Kankare J., Kulmala S., Anal. Chim. Acta, 1985, 171:259
138 Kaapkaak K., Kankare J., Puhakka O.,Anal. Chim. Acta, 1988, 207:195
139 Kaapkaak K., Kankare J., Lipiainen K., Anal. Chim. Acta, 1988, 215:341
140 Sung Y. E., Gaillard F., Bard A. J., J. Phys. Chem., 1998, B102:9797
141 Sung Y. E., Bard A. J.,J. Phys. Chem. B, 1998, 102:9806
142 Chiang M. T., Whang C. W., J. Chromatogra. A, 2001, 934:59
143 Jackson W. A., Bobbit D. R., Microchem. J., 1994, 49:99
144 Qin W., Zhang Z., Liu H., Anal. Chem., 1998, 70(9): 3579
145 章竹君《陕西师范大学学报》2000,28(3):79
146 郑兴旺,章竹君《高等学校化学学报》1999,21(2):124
147 郑兴旺,章竹君《分析化学》1999,27(2):145
148 黄家初,张成孝,章竹君《分析化学》1999,27(1):41
149 Zhang C.X, Zhang Z. J.,Anal. Len., 1998, 31(11): 1917
150 Zheng X. W., Zhang Z. J., Analyst, 1999, 124:763
151 何品刚,刘祥萍,余惠,方禹之《分析化学》2000,28(9):1062
152 张棘,严风霞,王筱敏,方禹之《光谱学与光谱分析》1995,15(1):109
153 杨敏丽,刘传之,何品刚,方禹之《高等学校化学学报》2002,23(11):2063
154 Yang M. L., Liu C. Z., He. P. G., Fang Y. Z.,Anal. Chim. Acta, 2002, 461:141
155 Littig J. S., Nieman T. A., Anal. Chem., 1992, 64:1140
156 Lin J. M., Yamada M., Microchem.,J., 1998, 58:105
157 Chen G. N., Lin R. E., Duan J. P., Zhang L., Anal. Chim. Acta, 1997, 341:251
158 Kondo T., Ito Y., Nomura S., Uosaki K., Thin Solid Films, 1996, 285:652
159 Irons G. P., and Greenway G. M., Analyst, 1995, 120:477
160 Heinze K. F., and Nieman T. A., Anal. Chim. Acta, 1993, 284:227
161 王伦《分析测试学报》1995,25(5):47
162 王伦,方宾《理化检验-化学分册》1997,33(2):31
163 Atwater J. E., DeHart J., Wheeler Jr R. R., J. Biolum. Chemilum., 1998, 13:125
164 Ouyang C. S., Wang C. M., J. Electrochem. Soc., 1998, 145(8) : 2654
165 Ouyang C. S., Wang C. M., J. Electroanal. Chem., 1999, 474: 82
166 Qin. W., Zhang Z. J., Liu H. J., Anal Chim. Acta, 1997, 357: 127
167 Zhang G. R, Chen H. Y., Anal. Chim. Acta, 2000, 419: 25
168 Slaterbeck A. F., Ridgway T., Seliskar C. J., Heineman W. R., Anal. Chem., 1999, 71: 1196
169 Zhao C. Z., Egashira N., Kurauchi Y., Ohga K., Electrochim. Acta, 1998, 43: 2167
170 Zhang X., Bard A. J., J. Phys. Chem., 1988, 92: 5566
171 Sato Y, Uosaki K.,J. Electroanal. Chem., 1995, 384: 57
172 Collinson M., Novak B., Martin S. A., Taussig J. S., Anal. Chem., 2000, 72:2914
173 Conner B. J., Reyes C. M., Morin C., Itakuura K., Teplitz R. L., Wallace R. B., Proc. Natl. Acad Sci. USA., 1983, 80:278
174 Riordan J. R., Rommens J. M., Kerm B., Alon N., Science, 1989, 245: 1066
175 MacDonald M. E., Ambrose C. M., Duyao M. P., Myers R. H., Cell, 1993, 72: 971
176 Jacobs K., Wolf S. F., Haines L., Fisch J., Kremsky N., Gougherty J. P., Neucleic Acids Res., 1987, 15: 2911
177 Part P. O., Lopez E., Mathis G, Appl Biochem., 1991,195: 283
178 Hurskainen P., Dahlen P., Ylidoski L., Kwiatkowski M., Siitari H., Lovgren T., Nucleic Acid Res., 1991,19: 1057
179 McNeil J. A., Johson C. V., Carter K. C., Anal. Tech. Appl., 1991, 8: 41
180慈云祥,常文保,帖建科《分析化学》1992,20(9) :100
181 Arnold J. L. J., Hammond P. W., Weie. W. A., Neison. N. C., Clin. Chem., 1989, 35:1588
182 Gillevet P. M., Nature, 1990, 348: 657
183 Pollard-Knight D., Simmonds A. C., Schaap P. A., Akhavan H., Brady M. A. W, Anal. Biochem., 1990, 185: 353
184 DeFillipo K A., Grayeski M. L., Anal. Chim. Acta, 1991, 249: 155
185 Wang. J., Cai X., Rivas G, Shiraishi H., Anal. Chim. Acta, 1996, 326:141
186 Wang J., Rivas G., Cai. X., Anal. him. Acta, 1997, 337:41
187 黄海珍,杨秀荣《分析化学》2002,30(4):491
188 杨丽菊,彭图治《分析化学》2001,29(3):355
189 赵湛,崔大付《传感器》2000,(8):63
190 黄智伟,黄琛《传感器技术》2001,20(1):4
191 高志贤,晁福寰《生物技术通讯》2000,11(1):45
192 胡伟平,张先恩,张治平《生物工程进展》1997,17(5):23
193 林琳,江龙《化学通报》2001,(5):261
194 陈帆,刘盛辉《浙江工业大学学报》1999,27(4):329
195 Wang J., Anal. Chim. Acta, 2002, 469:63
196 叶为全,周康源,王君等《传感器技术》2000,19(4):7
197 秦自楷《压电石英晶体》[M]北京:国防工业出版社,1980,68-112
198 Fawcett N. C., Evans J. A., Chien L. T., Anal. Lett., 1988, 21 (7): 1099
199 Okahata Y., Matsunobu Y., Ijiro K., et al., J. Am. Chem. Soc., 1992, 114:8299
200 Niikura K., Matsuno H., Okahata Y., J. Am. Chem. Soc., 1998, 120:8537
201 Caruso F., Rodda E., Furlong D., et al., Anal. Chem., 1997, 69:2043
202 赵秋红,林琳,唐季安,段明星,江龙,《科学通报》2001,46(4):292
203 Piunno P. A. E., Krull V. J., Hudson R. H. E., et al., Anal. Chim. Acta, 1994, 288: 205
204 Kleinjung F., Bier F. F., Warsilnke A., et al, Anal. Chim. Acta, 1997, 350:51
205 Graham C. R., Leslie D., Squirrell D. J., Biosens. Bioelectron., 1992, 7; 487
206 周建军 生物芯片[J]《国外医学分子生物学分册》2000,22(1):5
207 Ferguson J. A., Boles T. C., Adams C. P., et al., Nat. Biotechnol., 1996, 14(13):1681
208 Healey B. G., Matson R. S., Walt D. R., Anal. Biochem., 1997, 251: 270
209 姜广奋,陈润生,阎宏,欧阳颀,《中国科学》2000,30(5):461
210 Bier F. F., Kleinjang F., Scheller F. W., Sens. and Actuators B, 1997, 38:78
211 Thiel A. J., Frutos A. G., Jordan C. E., et al., Anal. Chem., 1997, 69:4948
212 Watts H. J., Yeung D., Parkes H., Anal. Chem., 1995, 67:4283
213 Pividori M. I., Merkoci A., Alegret S., Biosens. Bioelectron., 2000, 15: 291
214 Wang J., Cai. X., Jonsson C., Balakrishnan M., Electroanalysis, 1996, 8: 20
215 Wang J., Cai. X., Rivas G, Shiraishi H., Anal. Chim. Acta, 1996, 326: 141
216 Wang J., Cai. X., Rivas G, Shiraishi H., Farias P. A. M., Dontha N., Anal. Chem., 1996, 68: 2629
217 Wang J., Cai. X., Fernandes J. R., Grant D. H., Ozsoz M., Anal. Chem., 1997, 69:4056
218 Wang J., Cai. X., Rivas G, Shiraishi H., Farias P. A. M., Dontha N., Biosens. Bioelectron., 1997, 12: 587
219 Marrazza G, Chianella L., Mascini M., Biosens. Bioelectron., 1999, 14: 43
220 Hashimoto K., Ito K., Ishimori Y, Anal. Chim. Acta, 1994,286: 219
221 Li. L., Wang M., Dong S., Wang E., Anal. Sci. 1997, 13: 305
222 Pang D-W, Abruna H. D.,Anal. Chem., 1998, 70: 3162
223 Palecek E., Electroanalysis, 1996, 8: 7
224 Millan K. M., Spurmanis A. L., Mikkelsen S. K., Electroanalysis, 1992, 4: 929
225 Millan K. M., Mikkelsen S. K., Anal. Chem., 1993, 65: 2317
226 Millan, K. M., Saraullo A., Mikkelsen S. K., Anal. Chem., 1994, 66: 2943
227 Mikkelsen S. K., Electroanalysis, 1996, 8: 15
228 Liu S., Ye L., He P., Fang Y, Anal. Chim. Acta., 1996, 335: 239
229 Sun X., He. P., Liu S., Fang Y, Talanta, 1998,47: 487
230 Stell A. R., Herne T. M., Tarlov M. J., Anal. Chem., 1998, 70: 4670
231 Moser I., Schalkhammer T., Pittner R., Urban G, Biosens. Bioelectron., 1997, 12:729
232 Herne T. M., Tarlov M. J., J. Am. Chem. Soc., 1997,119(38) : 8916
233 Hashimoto K., Ito K., Ishimori Y, Anal. Chem., 1994, 66; 3830
234 Zhao Y D., Pang D. W, Hu S., et al., Anal. Chim. Acta, 1999, 388: 93
235 Wang J., Rivas G, Cai X., Electroanalysis, 1997, 9: 395
236 Wang J., Rivas G, Cai X., et al., Anal. Chim. Acta, 1993, 344: 111
237 Wang J., Rivas G, Cai X., et al., Anal. Chim. Acta, 1997, 347: 1
238 Wang J., Rivas G, Fernandes J. R., et al., Anal. Chim. Acta, 1998, 375: 197
239 Wang J., Palecek E., Nielsen P. E., et al., J. Am. Chem. Soc., 1996, 118: 7667
240 Palanti S., Marrazza G, Mascini M., Anal. Lett., 1996,29(13) : 2309
241 Arzum E., Kagan S., Burcu M., Mehmet O., Electroanalysis., 2001, 13: 219
242刘盛辉,何品刚, 方禹之《分析化学》1996,24911) :1301
243 Tanious F. A., Yen S-F, Wilson W. D., Biochemistry, 1991, 30: 1813
244 Takenaka S., Uto Y., Saita H., Yokoyama M., Kondo H., Wilson W. D., Chem. Commun., 1998,1111
245 Takenaka S., Takagi M., Bull. Chem. Soc. Jap., 1999, 72(3) : 327
246 Takenaka S., Takagi M., Uto Y, Kondo H., Nucleic Acids Symposium Series, 1998,39:107
247 Carrondo M., Coll M., Aymami J., Wang A., Marel G, Boom J., Rich A., Biochemistry, 1989,28: 7849
248 Takenaka S., UtoY, Kondo H., Ihara T., Takagi M., Anal. Biochem., 1994,218: 436
249 Xu C., Cai H., He P.G, Fang Y.Z., Fresen'. J. Anal. Chem., 2000, 367,593: 119.
250 Xu C., Cai H., He P.G, Fang Y.Z., The Analyst, 2001,126: 62
251 Xu C., He P.G, Fang Y.Z.. Anal. Chim. Acta., 2000,411:31
252 Cai H, Xu Y, He P. G, Fang Y. Z., J. Electroanal. Chem., 2001, 510: 78
253 Cai H, Xu Y, Zhu N. N., He P. G, Fang Y. Z.,The Analyst. 2002,127: 803
254 Cai H, Wang Y. Q., He P. G, Fang Y. Z Anal. Chim. Acta, 2002, 469: 165
255蔡宏,王延琴,何品刚,方禹之《高等学校化学学报》 已录用.
256 Lumley T. De., Campbell C., Heller A., J. Am. Chem. Soc., 1996,118: 5504
257 Patoisky F., Katz E., Bardea A., Willner I., Langmuir, 1999,15: 3703
258 Cardosi M. F., Standley C. J., Turner A. P. F., abstract, Second International Meeting on Chemical Sensors, Bordeaux, France
259 Wang J., Nasser A., Erdem A., Salazare M., Analyst, in press
260 Ropp P., Thorp H. H., Chem. Biol, 1999,6: 599
261 Napier M. E., Mikulecky K., Scaboo K., Eckhardt A., Baron N., Popovich N., Geladi M., Clin. Chem., 2000,46: A207
262 Kelley S., Boon E. M, Barton J. K., Jackson M., Mill M., Nucleic Acids Res., 1999,27: 4830
263 Boon E. M., Ceres D., Drummond T., Hill M., Barton J. K., Nat. Biotech., 2001, 18:1096
264 Korri-Youssoufi H., Garnier F., Srivtava P., Godillot P., Yassar A., J. Am. Chem. Aoc., 1997, 119:7388
265 Wang J., Jiang M., Fortes A., Mukherjee B., Anal. Chim. Acta, 1999, 402: 7
266 Siontorou C., Nikolelis D., Piunno P., Krull U., Electroanalysis, 1997, 9: 1067
267 Hianik J., Fajkus M., Sivak B., Rosenberg I., Kois P., Wang J., Electroanalysis, 2000,12: 495
268 Aoki H., Bulhmann P., Umezawa Y, Electroanalysis, 2000,12: 1272
269 Wang J., Rivas G., Cai X., et al., Anal. Chim. Acta, 1997, 337:41
270 Hashimoto K., Ito K., Ishimori Y, Sens and Actuators B, 1998, 46: 220
71 吴少惠,陈誉华,刘军等《中国公共卫生》2000,16:429
272 Tombelli S., Mascini K. M., Brassini L., et al., Biosensors and Bioelectron., 2000, 15: 363
273 Brabec V., Electrochim. Acta, 2000, 45: 2929
274 Maeda M., Anal. Sci., 1992, 8(1) : 83