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1. 发明授权

US08263411B2 Capacitive morpholino diagnostics for analysis of nucleic acids 有权
标题翻译：用于分析核酸的电容性吗啉诊断
公开(公告)号：US08263411B2
公开(公告)日：2012-09-11
申请号：US12362071
申请日：2009-01-29
申请人： Rastislav Levicky , Napoleon Tercero , Kang Wang , Ping Gong , Kenneth Shepard
发明人： Rastislav Levicky , Napoleon Tercero , Kang Wang , Ping Gong , Kenneth Shepard
IPC分类号： G01N33/50
CPC分类号： C12Q1/6825 , Y10T436/143333 , C12Q2525/113
摘要： A method to monitor the progress of hybridization between nucleic acid strands in solution and Morpholino strands immobilized on a solid support such as a working electrode in-situ, in real-time, and using label-free electrochemical measurements sensitive to hybridization-induced changes in the near-surface dielectric constant and charge organization.
摘要翻译：监测溶液中核酸链之间杂交进程的方法和实时固定在固体支持物如工作电极上的Morpholino链，并使用对杂交诱导的变化敏感的无标记电化学测量近表面介电常数和电荷组织。

2. 发明申请

US20100187133A1 CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS 有权
标题翻译：用于核酸分析的电容性多元醇诊断
公开(公告)号：US20100187133A1
公开(公告)日：2010-07-29
申请号：US12362071
申请日：2009-01-29
申请人： Rastislav Levicky , Napoleon Tercero , Kang Wang , Ping Gong , Kenneth Shepard
发明人： Rastislav Levicky , Napoleon Tercero , Kang Wang , Ping Gong , Kenneth Shepard
IPC分类号： G01N27/26
CPC分类号： C12Q1/6825 , Y10T436/143333 , C12Q2525/113
摘要： Surface hybridization, a reaction in which nucleic acid molecules in solution react with nucleic acid partners immobilized on a surface, is widely practiced in life science research. In these applications the immobilized partner, or “probe”, is typically single-stranded DNA. Because DNA is strongly charged, high salt conditions are required to enable binding between analyte nucleic acids (“targets”) in solution and the DNA probes. High salt, however, compromises prospects for label-free monitoring or control of the hybridization reaction through surface electric fields, as well as stabilizes secondary structure in target species that can interfere with probe-target recognition. In this work, initial steps toward addressing these challenges are taken by introducing Morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of Morpholino probes on gold supports can be fabricated with methods similar to those employed with DNA, and are shown to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for Morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Interestingly, positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a near-linear relationship, and demonstrates sensitivities (limits of quantification) in the pg mm−2 range.
摘要翻译：表面杂交，其中溶液中的核酸分子与固定在表面上的核酸配偶体反应的反应在生命科学研究中广泛应用。在这些应用中，固定化的配偶体或“探针”通常是单链DNA。由于DNA是强电荷的，因此需要高盐条件才能使溶液中的分析物核酸（“靶”）与DNA探针结合。然而，高盐不利于通过表面电场进行无标记监测或控制杂交反应的前景，以及稳定可能干扰探针 - 目标识别的靶物种中的二级结构。在这项工作中，通过引入Morpholinos（一类不带电荷的DNA类似物）进行表面杂交应用来采取解决这些挑战的最初步骤。可以用与DNA一起使用的方法类似的方法制备金载体上的Morpholino探针的单层，并且显示与靶链有效地并且序列特异性地杂交。用电化学方法分析杂交诱导的界面电荷组织变化，并比较Morpholino和DNA探针单层。通过与数值泊松 - 玻尔兹曼计算进行比较，可以确定和解释连接表面杂交态与界面电容的分子机制。有趣的是，取决于由施加电位控制的移动离子的表面种群，杂交的阳性和负电容反应（对比反转）是可能的。表面电容与目标覆盖（目标/面积）的定量比较显示近似线性关系，并显示pg mm-2范围内的灵敏度（定量限度）。

3. 发明申请

US20120211375A1 CAPACITIVE MORPHOLINO DIAGNOSTICS FOR ANALYSIS OF NUCLEIC ACIDS 有权
标题翻译：用于核酸分析的电容性多元醇诊断
公开(公告)号：US20120211375A1
公开(公告)日：2012-08-23
申请号：US13423228
申请日：2012-03-18
申请人： Rastislav Levicky , Napoleon Tercero , Kang Wang , Ping Gong , Kenneth Shepard
发明人： Rastislav Levicky , Napoleon Tercero , Kang Wang , Ping Gong , Kenneth Shepard
IPC分类号： G01N33/50 , G01N27/26
CPC分类号： C12Q1/6825 , Y10T436/143333 , C12Q2525/113
摘要： Use of Morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of Morpholino probes on gold supports are fabricated with methods similar to those employed with DNA, and are used to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for Morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a near-linear relationship, and demonstrates sensitivities (limits of quantification) in the pg mm−2 range.
摘要翻译：使用Morpholinos，一类不带电荷的DNA类似物，用于表面杂交应用。使用类似于DNA使用的方法制备金支持物上的Morpholino探针的单层，并且用于与靶链有效地和序列特异性地杂交。用电化学方法分析杂交诱导的界面电荷组织变化，并比较Morpholino和DNA探针单层。通过与数值泊松 - 玻尔兹曼计算进行比较，可以确定和解释连接表面杂交态与界面电容的分子机制。取决于由施加电位控制的移动离子的表面种群，对于杂交，阳性以及负电容反应（对比反转）是可能的。表面电容与目标覆盖（目标/面积）的定量比较显示近似线性关系，并显示pg mm-2范围内的灵敏度（定量限度）。

4. 发明申请

US20100035248A1 SURFACE-BASED NUCLEIC ACID ASSAYS EMPLOYING MORPHOLINOS 审中-公开
标题翻译：基于表面的核酸测定使用莫比诺
公开(公告)号：US20100035248A1
公开(公告)日：2010-02-11
申请号：US12139477
申请日：2008-06-15
申请人： Rastislav Levicky , Napoleon Tercero , Ping Gong , Kenneth Shepard
发明人： Rastislav Levicky , Napoleon Tercero , Ping Gong , Kenneth Shepard
IPC分类号： C12Q1/68 , C12M1/34
CPC分类号： C12Q1/6834 , C12Q2525/113
摘要： The sequence determination, detection, and quantification of nucleic acid molecules through sequence-specific binding (hybridization) on a solid support, specifically when Morpholinos are used as the surface-immobilized probe species in surface-based nucleic acid assays, and the assays as disclosed herein.
摘要翻译：通过在固体支持物上的序列特异性结合（杂交），特别是当在基于表面的核酸测定中将Morpholinos用作表面固定化探针物质时，核酸分子的序列测定，检测和定量以及所公开的测定这里。

5. 发明授权

US08273578B2 Capacitive morpholino diagnostics for analysis of nucleic acids 有权
标题翻译：用于分析核酸的电容性吗啉诊断
公开(公告)号：US08273578B2
公开(公告)日：2012-09-25
申请号：US13423228
申请日：2012-03-18
申请人： Rastislav Levicky , Napoleon Tercero , Kang Wong , Ping Gong , Kenneth Shepard
发明人： Rastislav Levicky , Napoleon Tercero , Kang Wong , Ping Gong , Kenneth Shepard
IPC分类号： G01N33/50
CPC分类号： C12Q1/6825 , Y10T436/143333 , C12Q2525/113
摘要： Use of Morpholinos, a class of uncharged DNA analogues, for surface-hybridization applications. Monolayers of Morpholino probes on gold supports are fabricated with methods similar to those employed with DNA, and are used to hybridize efficiently and sequence-specifically with target strands. Hybridization-induced changes in the interfacial charge organization are analyzed with electrochemical methods and compared for Morpholino and DNA probe monolayers. Molecular mechanisms connecting surface hybridization state to the interfacial capacitance are identified and interpreted through comparison to numerical Poisson-Boltzmann calculations. Positive as well as negative capacitive responses (contrast inversion) to hybridization are possible, depending on surface populations of mobile ions as controlled by the applied potential. Quantitative comparison of surface capacitance with target coverage (targets/area) reveals a near-linear relationship, and demonstrates sensitivities (limits of quantification) in the pg mm−2 range.
摘要翻译：使用Morpholinos，一类不带电荷的DNA类似物，用于表面杂交应用。使用类似于DNA使用的方法制备金支持物上的Morpholino探针的单层，并且用于与靶链有效地和序列特异性地杂交。用电化学方法分析杂交诱导的界面电荷组织变化，并比较Morpholino和DNA探针单层。通过与数值泊松 - 玻尔兹曼计算进行比较，可以确定和解释连接表面杂交态与界面电容的分子机制。取决于由施加电位控制的移动离子的表面种群，对于杂交，阳性以及负电容反应（对比反转）是可能的。表面电容与目标覆盖（目标/面积）的定量比较显示近似线性关系，并显示pg mm-2范围内的灵敏度（定量限度）。

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