CN101039750A - 对可通过导管进入测量室中的液态测量样品进行电化学测量的方法和相应的布置 - Google Patents
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Abstract
特别在一具有一微流体系统的封闭样品管中使用储存干燥试剂来进行所谓的酶联DNA杂交实验时，必须在测量之前即时将所述试剂溶解在所述微流体系统中，并将其输送至测量室。其中，必须避免在将所述试剂溶入水中时在液态试剂的前方产生不能进入所述测量室的气泡。因此，本发明对液态测量样品和液态试剂(R1，R2)的输送方式为，先使所述气泡进入废料导管(W1，W2)，再以不产生气泡的方式将所述测量样品和所述试剂(R1，R2)送入所述测量室(150)。借此可避免出现误测。
Description
技术领域
本发明涉及一种对可通过导管进入的测量室中的一液态测量样品进行电化学测量的方法，为进行电化学测量输送至少一种液态试剂。此外，本发明还涉及一种用于实施所述方法的相应布置和所述布置的应用。
背景技术
为了解答人类基因问题，核酸分析，例如对全血中的白细胞的分析，必须先在样品制备步骤第一阶段破碎所述的白细胞，随后将在此过程中释放出来的DNA隔离。第二阶段使用聚合酶链反应(Polymerase Chain Reaction，PCR)实现选择性的DNA复制(扩增)，以便将需要检测的DNA浓度提高，从而在第三阶段时可对其进行检测。
后面的几个子步骤在实验室中根据已知的现有技术分别地进行。上文所述的三个阶段分别包括两个以上的工作步骤，且所述三个阶段彼此分别地使用不同仪器实施。各工作步骤实质上为手动实施。
这些步骤的实现依赖于实验室仪器，例如细胞破碎仪；一PCR仪(即所谓的热循环仪)，可选的为一个适用于定量PCR的PCR仪；一电泳仪；一杂交站；一光阅读器，所谓的微量离心管(Eppendorf-Tubes)；两个以上的吸液器与一用于冷却试剂的冷却容器；并且必须由受过训练的人员在遵守有关感染危险、废料清理或诸如此类的安全规章的情况下实施。尤其是必须对试剂溶液进行两次以上的容积定量，即精确定量(吸液)。这些步骤不仅费时，且成本高昂。
现有技术中已知有用于生物化学分析的设备，其根据WO 02/073153特别使用基于硅的测量模块，这些测量模块可集成在一芯片中。其中，根据另一份WO 02/072262 A1中，在分析模块内以干燥存放的形式集成有用于分析所需的试剂。
发明内容
本发明的目标是在一小型样品管中实现一成本低廉、操作简单的完整DNA或蛋白质分析过程。从这一目标出发，本发明的目的是在一测量室中进行一电化学测量，并为此以不产生气泡的方式输送测量样品和所用的液态试剂，所述测量样品和液态试剂均以泵入方式被送入所述测量室内(本发明特别适用于这样一种分析，但本发明的用途并不仅限于此)。此外，本发明还提供一种用于实施所述方法的布置。
这个目的通过根据权利要求1所述的处理步骤序列而达成。权利要求11中讲述的是一种相应的布置。从属权利要求涉及的是所述方法和所述的相应装置的细化方案。
本发明涉及一种借助一相应布置而实现的方法，用于将液体输送至一测量室内进行电化学测量，所述液体特别为一液态样品和至少一种液态试剂，实施所述方法时，所有参与的液体内都不会出现气泡。在通过溶解固体试剂来制备液态试剂的情况下，这一点特别有意义。
液态样品和液态试剂存在于通向所述测量室的不同导管中，所述液态样品和液态试剂彼此之间被空气所隔离，并且所述液态样品和液态试剂与所述测量室之间也同样被空气隔离，通过本发明可以以不产生气泡的方式将所述液态样品和液态试剂输送到测量室内，并借此确保测量室内实际测量过程不受到干扰。
实施本发明时，从不同侧向所述测量室输送测量样品和试剂是有利的。在与此相关的布置中，所述测量室的不同侧存在用于排气的废料通道。
借助这种布置和本发明的方法可在测量之前对用于向测量室输送液态物质的导管进行排气。这一点在下述情况下特别重要，即，用于核酸诊断的样品管使用的是干燥试剂，在进行真正意义上的诊断或测量过程之前，所述干燥试剂在水中被直接“就地”溶解，形成一液态试剂，并被输送至测量室。这是因为在此过程中，通过主动泵入依次进入测量室的液态试剂和测量用液体的前方会不可避免地出现气泡。对于测量室而言不期望出现这种气泡，因为可能再也无法排出空气，并因而对电化学测量造成干扰或阻碍。
本发明特别应用在所述样品管中进行实际检测的子区域中。所述检测包括酶联DNA杂交检测。其中，用一合适的酶(例如与链霉亲和素结合的碱性磷酸酶)标示杂交结果，并通过测量一因酶活性而产生的产物(例如p-氨基酚)对杂交结果进行检测。但本发明也可用于其他液态样品的测量过程，该液态样品最初需要通过主动泵入与试剂溶液一起进入测量室，例如酶联免疫吸附实验(ELISA：Enzyme linked Immuno sorbed Assay)。
附图说明
下面借助附图所示的实施例并结合权利要求对本发明的其他特点和优点进行说明，其中：
图1为一具有一导管系统的样品管示意图，所述导管系统带有相应的功能标识；
图2为导管的示意图，所述导管具有用于储存干燥试剂的凹槽；
图3、4为图2所示的导管的横截面图，所述导管具有用于储存干燥试剂的凹槽；
图5为本发明布置的第一实施例示意图，其中，用于输送试剂和测量样品的导管布置在测量室的一侧；以及
图6为本发明布置的第二实施例示意图，其中，用于输送测量样品和试剂的导管布置在测量室的不同侧。
具体实施方式
相同的单元在各附图中用相同的参考符号表示。下文分别将图1至图4以及图5和图6结合在一起进行充分说明。
图1显示的是一具有一导管系统的样品管(Cartridge)100，所述导管系统具有复数个布置在所述样品管底板中的微型通道或空腔，和一个将所述样品管底板封闭的覆膜。样品管100具体而言由一塑料壳体101构成，所述塑料壳体带有规定结构的微流体系统，下文中将借助图2至图4对所述结构加以说明。
从图1所示的示意图中可以看到一样品入口102，其后连接一个定量段105。所述定量段后依次连接有一用于细胞破碎的通道区110和一用于进行PCR的区域120。通过阀122、122′可将实际的PCR室关闭。在区域130中特别根据酶联DNA杂交法进行样品检测。
从图1中还可看到入水口103至103。此外还存在排气口104至104。
通道系统中存在用于容纳废料的其他区域106、107、108、109。此外还存在一用于容纳试剂的区域131、131′。
图2至图4显示的是图1所示的试剂通道131、131′的布局和结构。如图3所示，凹槽132至1325′适用于容纳干燥试剂133至1335′。在图4中，凹槽132至1325′已充填有干燥试剂133至1335′。
图5和图6所示的参考符号150表示一用于进行电化学测量的测量室，所述电化学测量尤其为所谓的酶联DNA杂交实验。为进行这种测量，必须将一杂交测量样品和特定的试剂送入测量室。图5和图6没有显示测量构件和用于获取电信号的构件。
测量室在图5和图6中显示为椭圆形空腔150，并在相对的两侧具有入口151和152，所述入口构成通向导管的接口。测量室150通过入口151与废料通道W1相连。另一入口152以相同方式与废料导管W2相连。废料导管通过阀与外界相连。通过阀的开关可确定流体系统中的流动方向。所述阀的一特别功能是，其只能由空气通过，因而可阻止试剂和测量样品与外界接触。
借此可实施下述处理过程：通过一属于样品管100的外设泵将样品输送入测量室150，其中，液体前面可能存在的气泡被向前推进。由于测量样品的体积大于测量室的体积，因此，通过入口151或152分别产生了将气泡和测量样品推入废料导管W1或W2的推进力。
随后输送一第一试剂R1，使得气泡进入其中一个废料通道W1或W2，而并不进入测量室150。这一过程也称为“排气”。在此之后，通过对上文所述的阀进行开关操作，可使试剂流过测量室150。
输送第二试剂R2时实施相同的过程。
液态样品和液态试剂存在于通向所述测量室的不同导管中，所述液态样品和液态试剂彼此之间被空气所隔离，并且所述液态样品和液态试剂与所述测量室之间也同样被空气隔离，借此可以以不产生气泡的方式将存在于不同导管中的液态样品和液态试剂输送到测量室内，并确保测量室内的实际测量过程不受到干扰。
图6所示的布置与图5所示的布置的区别在于，样品导管161与用于输送试剂的导管162和162′布置在测量室150相对的两侧。除此之外，所述布置与图1所示的布置相一致。
通过所述方法和图1与图5所示的布置，或所述方法和图1与图6所示的布置，可进行所需求的排气过程。此外，借助所述布置可在不产生负压(吸力)的情况下达到使液体双向(正向泵入和反向泵入)流过测量室的目的。借此可改善测量室内的结合过程。
Claims (15)
1.一种对可通过导管进入测量室中的液态测量样品进行电化学测量的方法，其中，为进行电化学测量输送至少一种液态试剂，所述方法包括下列措施：
先通过一入口将所述测量样品泵入所述测量室，将所述测量样品的过量部分送入位于所述测量室与所述入口相对的另一侧的废料导管，
随后将所述的至少一种试剂从一导管输送入一位于所述测量室的相同侧的废料导管，所述试剂先不流过所述测量室，以及
再以与所述测量样品发生相互作用为目的，以不产生气泡的方式使所述试剂流过所述测量室并进入另一废料导管。
2.根据权利要求1所述的方法，其特征在于，
通过不同的入口将所述测量样品和所述的至少一种试剂输送入所述测量室。
3.根据权利要求1或2所述的方法，其特征在于，
将所述测量样品和所述的至少一种试剂前面的气泡输送入不同的废料导管。
4.根据上述权利要求中任一项权利要求所述的方法，其特征在于，
所述测量样品和所述试剂流过所述测量室后，所述测量室中不存在空气。
5.根据上述权利要求中任一项权利要求所述的方法，其特征在于，
使用两种试剂。
6.根据上述权利要求中任一项权利要求所述的方法，其特征在于，
通过在预定剂量的固体干燥试剂中添加一溶剂来“就地”形成液态试剂。
7.根据权利要求6所述的方法，其特征在于，
将水用作所述溶剂。
8.根据上述权利要求中任一项权利要求所述的方法，其特征在于，
所述方法用在进行过PCR的酶联DNA杂交检测上，为此使用一具有PCR室和标记-酶-试剂导管或酶-底板-试剂导管的样品管。
9.根据权利要求7所述的方法，其特征在于，
进行PCR时在所述试剂导管中充水。
10.根据权利要求8所述的方法，其特征在于，
所述杂交过程完成后对所述标记-酶-试剂导管或所述酶-底板-试剂导管进行排气，然后先用不带气泡的第一试剂对所述检测室进行冲洗，再用不带气泡的第二试剂对所述检测室进行冲洗，以便随后进行所述电化学测量。
11.一种用于实施根据权利要求1或根据权利要求2至10中任一项权利要求所述方法的布置，所述布置具有一测量室(150)，所述测量室具有至少两个入口(151，152)，其中，第一入口(151)与一用于容纳废料(Waste 1)的第一导管(W1)相连，第二入口(152)与用于容纳废料(Waste 2)的另一导管(W2)相连。
12.根据权利要求11所述的布置，其特征在于，
所述的其中一个入口(151)既构成所述测量样品的输送导管(161)的接口，又构成所述废料导管(W1或W2)的接口。
13.根据权利要求11或12所述的布置，其特征在于，
所述的另一入口(152)既构成用于输送至少一种试剂(R1，R2)的接口，又构成一废料导管(W1或W2)的接口。
14.根据权利要求11至13中任一项权利要求所述的布置，其特征在于，
存在用于输送两种试剂(R1，R2)的复数个导管(162，162′)。
15.根据权利要求11至14中任一项权利要求所述的布置，其特征在于，
所述用于输送试剂(R1，R2)的复数个导管(162，162′)和所述测量样品入口(161)布置在所述测量室(150)相对的两侧。
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| CNB2005800352122A Expired - Fee Related CN100534619C (zh) | 2004-10-15 | 2005-10-17 | 对可通过导管进入测量室中的液态测量样品进行电化学测量的方法和相应的布置 |
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| EP (2) | EP1807208B1 (zh) |
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| CN109536374A (zh) * | 2018-11-27 | 2019-03-29 | 南京先进激光技术研究院 | 一种具有避免气泡产生结构的试剂反应管 |
Also Published As
| Publication number | Publication date |
|---|---|
| EP1796838A1 (de) | 2007-06-20 |
| WO2006042838A1 (de) | 2006-04-27 |
| EP1807208B1 (de) | 2013-03-20 |
| US7851227B2 (en) | 2010-12-14 |
| JP2008517259A (ja) | 2008-05-22 |
| EP1807208A1 (de) | 2007-07-18 |
| US20090130658A1 (en) | 2009-05-21 |
| US20090136922A1 (en) | 2009-05-28 |
| WO2006042734A1 (de) | 2006-04-27 |
| JP4546534B2 (ja) | 2010-09-15 |
| EP1796838B1 (de) | 2014-10-08 |
| CN100534619C (zh) | 2009-09-02 |
| CN101039751A (zh) | 2007-09-19 |
| CN101039751B (zh) | 2010-05-05 |
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