CN100501387C - 用于进行横向流动分析的读数装置与方法和系统 - Google Patents
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Abstract
披露了用于进行横向流动分析的反光试剂盒,它包括读数装置(65)和膜式检测条(76)。所述读数装置(65)是便携式的。可以对体液进行分析,以便以高灵敏度检测某些激素,葡萄糖,或其他所关心的体液的存在。所述膜式检测条(76)可以容纳含有要检测的分析物的测试流体或测试样品。可以将所述膜式检测条直接插入读数装置(65)的容纳口。通过位于膜式检测条(76)上方的吸光部件(80)遮挡来自所述容纳口的散射光线,可以提高灵敏度,并且减少进入读数装置(65)的散射光线或环境光线。读数装置(65)还包括一个或多个能够检测反射电磁辐射(93)的强度的传感器(92)。
Description
技术领域
膜式检测装置,特别是在诊断药品中使用的装置,采用了多种内部和外部校准器,以便提供有关测试溶液中所关心的分析物的定性或定量结果。一种类型的膜式测试装置是横向流动分析装置。
背景技术
一般,横向流动分析装置是膜式检测装置,其中,将被怀疑含有所关心的分析物的样品在或靠近膜检测条的一端放置。由毛细作用通过所述膜式条的液相,将所述样品携带到所述膜条的相反一端。在通过所述膜条时,检测样品中的分析物(如果有的话)会遇到一种或多种“俘获”试剂,它可以与所述试剂起反应,以便产生可检测的信号。
诸如妊娠测试等的家用分析装置现在已经完善。家用分析装置可用于检测人体的生理学变化,其目的是改善个人的卫生和健康。消费者的健康意识越来越强,并且,如果消费者能够监测包括激素水平等在内的他或她自身的身体功能的话,将是非常有利的。
存在多种不同的分析方法,可以指示人体的生理学变化。另外,存在多种不同的分析装置,这些装置是通过读取分析条或检测样品而工作的。某些装置使用荧光发射,而其他装置使用光反射。
授予Catt等的美国专利号6235241B1(“Catt专利”)涉及与分析装置组合使用的分析结果读数器。类似于Catt专利中所披露的装置的市售装置被称为UNIPATH CLEAR PLAN 生殖力监测器。在本文的图1中示出了该装置,它包括生殖力监测装置21,具有一个可除去的手持盖子22,它结合在外壳25上的容纳装置23中。将体液涂在检测条24上,并且可以将检测条24放入容纳装置23,在这里,检测条24接收通过窗口26照射在检测条24上的光线。然后,分析反射光线水平,以便提供结果。
所述生殖力监测装置的一个问题是,在很多场合下,它们不能够提供高度的灵敏性。就是说,出于医学目的,需要监测某些分析物,但是,不需要高度的灵敏性或复杂的仪器来准确和精确地检测分析物的含量。很多现有的家用读数装置具有低的信噪比,这种信噪可能在某种程度上是由于将过量的散射光线或环境光线意外导入观察窗口而引起的。在利用反光型方案进行精确测定时,重要的是减少或消除散射环境光线量,以便获得高度的灵敏性。因此,非常需要提高信噪比,并且提高所述读数装置的灵敏度。
另一种家用读数装置被称为血糖计,它是由Indiana46250的Indiana polis的Boehringer MannheimDiagnostics公司生产和销售的。装置是为了在家中检测血糖含量而设计的反光型仪器。该仪器不采用横向流动分析。相反,要求用户将一滴血放置在检测垫上。该仪器的反光传感器部分包括一个可除去的支架,它具有两个矩形窗口。
本行业需要一种为横向流动分析检测条而设计的灵敏的读数装置。需要这样一种读数装置,它提供了用于快速将检测条放入到位以便接受读数或结果,同时避免过多环境光线和散射光线进入的有效并且可靠的装置。需要一种用于检测激素等的能提供高灵敏度的读数装置。具有获得高度光透射和反射效率的窗口的读数装置是有用的。
发明内容
一种改进的横向流动分析读数装置,该读数装置被设计成能够从横向流动分析膜条中检测分析结果,所述分析结果是通过检测分析物在所述膜条的检测区内的结合显现的,该装置包括:
(a)具有外部和内部的外壳;
(b)所述外壳内的容纳口,用于将膜条直接从所述外壳的外部纳入所述读数装置外壳的内部,所述容纳口具有一个遮光结构;
(c)读数机构,该机构包括:
(i)电磁辐射源;
(ii)一个或多个能够检测所述电磁辐射强度的传感器,其中,所述电磁辐射源和传感器位于所述读数机构内,以便当所述膜条被纳入所述容纳口时,并且由所述遮光结构封闭时,所述电磁辐射首先照射在所述膜条上的检测区,然后再照射在所述传感器上,其中该电磁辐射在进入容纳口之前通过一孔,该孔的尺寸大致为检测区的尺寸。
其中,所述容纳口包括压在所述膜条上的压力板。
其中,所述压力板是弹簧加载的。
在所述容纳口内提供吸光部件,以便吸收散射光线。
所述吸光部件包括位于容纳口内的膜条附近的吸收垫。
所述吸光部件包括柔性的和顺应性的毛毡材料。
所述电磁辐射首先通过孔,然后再进入所述容纳口。
所述孔是长形的。
其中,所述孔是圆形的。
其中,所述容纳口包括用于参考读数的第一个停止位置,和用于样品读数的第二个停止位置。
一种检测盒,包括权利要求1-10中任一项的横向流动分析读数装置,以及一个分析装置,它包括多孔液体可渗透的膜条,其中,所述多孔液体可渗透膜条的厚度是电磁辐射能够穿透的,其中,所述分析结果是通过可检测分析物与固定在所述检测区上的结合试剂的直接或间接地特殊结合显现的。
所述读数机构部分地通过用传感器检测从以下区域反射的电磁辐射来确定所述分析结果:
a)所述膜条的检测区;和
b)所述膜条上的校正区,
其中,对在a)和b)中测定的值进行比较。
一种用于进行横向流动分析,以便检测存在于检测液体中分析物的量的系统,该系统包括:
(a)能够产生可检测信号的探针-分析物结合物;
(b)膜条,所述膜条包括检测区,所述检测区具有沉积在它上面的第一种捕获试剂,其中,所述第一种捕获试剂被设计成用于结合探针-分析物结合物,以便固定所述探针-分析物结合物,从而在所述检测区内形成夹心复合物;和
(c)根据权利要求1-10中任一项的读数装置。
一种用于进行横向流动分析以便检测在检测液体中分析物的数量的方法,该方法包括:
(a)在膜条上提供探针-结合物,将所述探针-结合物设计成能产生可检测信号;
(b)在所述膜条上提供分析物;
(c)将所述探针-结合物结合在所述分析物上,以便形成探针-分析物结合物复合物;
(d)其中,将所述膜条设计成将含有探针和探针-分析物结合物复合物的检测液体固定,所述膜条包括检测区;
(e)在所述检测区提供第一种捕获试剂,其中,所述第一种捕获试剂是固定的,并且被设计成能结合所述探针-分析物结合物复合物,以便通过在所述检测区内形成夹心复合物,固定所述探针-分析物结合物复合物;
(f)提供读数装置,将所述读数装置设计成检测来自所述膜条的分析结果,所述读数装置包括具有外部和内部,以及具有遮光结构的容纳口的外壳,所述容纳口结合在所述外壳内,用于将所述膜直接从读数装置外壳外部纳入读数装置外壳内部;
(g)阻止环境光线到达所述膜检测区,以便提高灵敏度;
(h)其中,所述电磁辐射源和传感器是这样定位的,使得当所述膜条被纳入所述容纳口时,电磁辐射首先照射在所述膜条上的检测区,然后到所述传感器;
(i)设置使电磁辐射在进入容纳口之前通过一孔,所述孔的尺寸大致为检测区的尺寸。
还包括在容纳口中设置的一吸光部件以吸收散射光,该吸光部件包括吸收垫,该吸收垫靠近在容纳口内的膜条并且在使用中位于膜条的上表面上,该吸收垫完全覆盖这样的区域:在该区域下面该膜条被来自该膜条下面的光照射。
在本发明中,可以提供用于横向流动分析的读数装置,和用于进行分析的系统。将所述读数装置设计成用于检测来自膜条的分析结果,其中,所述结果是通过将可检测分析物固定在膜条的检测区内而显现的。所述分析读数装置包括外壳和位于该外壳内的容纳口。所述容纳口可以包括一个遮光结构,并且能接纳直接来自所述外壳外部的膜条。就是说,将膜条插入所述容纳口。可以将所述容纳口设计成减少导入所述读数装置的散射光线或环境光线。
还可以提供一种读数机构,它包括电磁辐射源,和一个或多个能够检测反射的电磁辐射的强度的传感器。可以将所述辐射源和传感器安装在所述读数机构内,以便当所述膜条被纳入所述容纳口时,所述辐射在照射所述传感器之前,先照射所述膜条上的检测区。
在本发明的另一种实施方案中,可以提供一种检测盒,它包括横向流动分析读数装置,和多孔的液体可渗透的膜条。
在本发明的另一种实施方案中,可以提供一种用于进行横向流动分析的系统,以便检测存在于检测液体中的分析物的数量。所述系统可以包括为了产生可检测信号而设计的探头,和为了移动检测液体而设计的膜条。所述膜条包括检测区。另外,采用了以前所披露的读数装置,它具有为了减少进入该读数装置的散射光线而设计的容纳口和遮光结构。通过本发明获得了具有较高灵敏度的分析结果。
附图说明
在本说明书中提供了本发明的完整的和可操作的说明,包括为本领域普通技术人员而提供的最佳方式。以下附图说明了本发明:
图2是本发明读数装置的一种实施方案的透视图,表示遮光结构和容纳口;
图3表示所述读数装置的透视图,其中,容纳口45业已向上进行了分解,以便显示细节;
图3a是顶板的底部视图,表示压力板与容纳口中的顶板的相互作用;
图4表示沿图2中线4-4的本发明一种实施方案的容纳口的横剖视图;
图5表示本发明读数装置的另一种实施方案,它具有位于该读数装置上表面上的通道,该通道被设计成用于容纳膜检测条;
图5a表示沿图5中线5a-5a的所述容纳装置的膜条容纳口的横剖视图;
图5b表示所述读数装置的电子部件的设计布局,包括微控制器,LCD显示器等;
图6表示在前面的图5中所示出的实施方案的膜条容纳孔的更近的示意图,表示一种特定的用途,其中,所述膜条包括连接到一个或多个切口中的小块;和
图7表示沿图6中线7-7的截取如图6所示的结构的横剖视图。
本发明的详细说明
现在参见本发明的实施方案,下面提供了它的一种或多种实施例。所提供的每一个实施例是为了说明本发明的,而不是作为对本发明的限制。事实上,本领域技术人员可以理解的是,在不超出本发明范围或构思的前提下,可以对本发明进行各种改进和改变。例如,作为一种实施方案一部分而示出或说明的特征,可以用在另一种实施方案上,以便得到另一种实施方案。因此,本发明意在包括属于所附权利要求书及其等同方案范围的这样的改进和改变。
在本发明中,提供了反光计或读数装置。所述读数装置可用于横向流动分析,以便提供定量结果。可以设计所述计量装置,以便提供改善了的灵敏度和提高了的精确度。本发明的方法和系统可用作更精确和灵敏的替代方案,以代替直接目测膜式分析条。
本发明的读数装置可以包括各种部件,包括光源,如发光二极管(LED)或激光,光束调制器,反光镜,透镜,光电二极管,样品支架和其他选择性部件,正如本文进一步说明的。在任何场合下,所述样品支架都有利于插入膜检测条,只有少量的环境光线或散射光线通过,从而降低了噪音水平。提供了一种具有较高灵敏度的,具有改善了的信噪比的读数装置。所述样品支架可以包括具有弹簧加载部件的机械设计。在某些用途中,为同一个膜检测条提供了至少两个不同的停止位置,其中,第一个停止位置可用于提供参考读数,而第二个停止位置可用于在检测部位或检测区对实际样品进行读数。
在图2中进一步示出了本发明的一种实施方案,其中,读数装置40将膜条41纳入容纳口45,以便提供结果。还示出了遮光结构28。膜条41上的检测区42距离参考区43一定距离,由参考区43提供基线参考值或校正读数。在所示出的具体实施方案中,所提供的检测区42朝向外侧,而参考区43朝向内侧,不过,应当理解的是,所述相应区域的位置可以与图2所示相反。
读数装置40可以包括壳体外部44,和开/关转换器49,和壳体内部(在图2中未示出)。在图2中,示出了LCD显示器60。
在图3中,示出了遮光结构28的一种示意图,其部件是从读数装置40的壳体外部44向上分解的。还示出了顶板50。在图3中示出的装置,相当于在图2中示出的装置,并且是大体上相同的实施方案。由底板56在底部边缘上限定了容纳口45的边界,并且在顶部边缘由顶板50限定边界。在容纳口45内,有一个压力板51,将膜条41插入它下面。通过弹簧52保持压力板51与膜条41的弹性接合(在图3中未示出)。将膜条41保持在孔54上面,在图3中,该孔是圆形的。不过,该孔可以具有很多不同的形状和尺寸,最优选的是接近要检验的膜条41上所关心的区域的尺寸和/或形状。通道53构成了插入膜条41的回路。由螺钉55a-d将顶板50向下保持在外壳44上。
在图3a中,示出了顶板50的底部,显示凹槽58。压力板51被放置在凹槽58内,通过弹簧52以弹力接合的形式固定。还示出了吸光部件57,它放置在膜条41的顶部或上表面(参见图2)。吸光部件57起着与孔54接触的低反光样品的作用,使得能够对该仪器进行校正,以便消除在传感器外壳内的内部反射的影响。在实践中,当首先给所述仪器提供能量时,可以通过微处理器自动进行所述校正。另外,吸光部件57可以吸收完全透过膜条41的所有光线,以便所述光线不会向下往回反射到传感器92上(参见图5a)。这样,提高了读数装置40的灵敏度和信噪比。
吸光部件57可以包括几乎任何类型的能够吸收光线的材料,如黑色或暗色绒屑,塑料,金属,毛毡或其他材料。例如,可以采用被用于照相技术的、已知能吸收光线的材料。所述材料可以是柔性的和/或顺应性的,并且可以包括毛毡。对于吸光部件57来说,没有优选的具体尺寸或形状。不过,重要的是,吸光部件57完全覆盖这样的区域,在所述区域下面膜条41受到来自其下部的光线照射的。吸光部件57的一种选择性特征是通过采用毛毡或可悬垂的材料提供适于检测条的柔性或顺应性形式。
图4表示遮光结构28的横剖视图,具有如图2中的线4-4所示的容纳口45。容纳口45包括压力板51,它结合在顶板50和底板56之间。将膜条41插入压力板51下面,其中,膜条41的检测区42可直接放置在光线通道59上面。由诸如发光二极管(LED)的光源(现在如图4所示)产生的光线,沿箭头59a方向向上通过,并且沿图4所示箭头59b方向从膜条41上向下反射。
图2-4所示出的读数装置的内部发光和传感部件与图5-5a所示装置基本上相同。
对读数装置40的灵敏度重要的是,直接位于膜条41下面的光照孔的大小,接近膜条41上的检测区42的大小。在其他用途中,所述孔(在图4中未示出)可以略大于检测区42。在某些场合下,所述孔可以比检测区42的面积大大约1.3倍或甚至1.8倍。不过,业已发现,所述孔的尺寸越接近膜条41上检测区42的尺寸,通过读数装置40获得的信噪比越高,并且读数装置40也越灵敏。另外,膜条41还可以包括位于膜条41上的另一个位置处的参考区43。参考区43可以位于光线通道59上面,以便获得读数装置40的参考读数或校正读数。然后,在第二个步骤中,可以将检测区42放置在光线通道59上面,以便获得样品读数。以剖面形式示出了位于吸光部件57上面的弹簧52,它正好结合在膜条41上面。吸光部件57能够吸收从外面意外进入容纳口45的光线。另外,吸光部件57能够吸收可以通过光线通道59,并且完全透过膜条41的光线。这样防止了散射光线的反射,提高了灵敏度。
在图5中示出了本发明的另一种实施方案。在LCD显示器74下面提供了一个遮光结构81。该遮光结构是从上面通过顶板72限定的,并且从下面通过底板78限定。读数装置65包括外壳73,它具有在顶部由罩66界定的容纳口64。容纳口64的一部分包括通道68,如图5所示,该通道是垂直分布的。孔69(在图5中该孔是矩形形状的)位于通道68的底部。作为定位点,提供了第一切口70和第二切口71,以便容纳具有小块77的膜条,可以从图5a中看出这一点。螺钉67a和67b将罩66向下固定在顶部72上。罩66的作用是减少照射在孔69附近的环境光线的量,提高读数装置65的灵敏度,并且提高所获得的结果的信噪比。示出了靠近外壳73右侧的开/关转换器75。
图5a是沿图5的横剖线5a-5a的基本示意图,表示在本发明中所采用的读数装置65的基本内部结构。由螺钉67a-b将顶板72向下固定在底板78上,并且还起着将罩66固定在板72上的作用。在横剖面上,人们可以看到位于膜条76上面的吸光部件80。小块77结合在第一切口70中,以便使膜条76位于合适的位置中,以便接收来自发光二极管(LED)90的光线91。光线91通过膜条76,然后沿光线通道93向下反射到传感器92上。在某些用途中,传感器92是二极管。还可以看到外壳73,并且可以包括在图5a中未示出的其他部件。
在图5b中示出了读数装置65的基本示意图。在图5b中,在图5b的右侧示出了具有16色的LCD显示器74。LCD显示器74与微控制器95连接。微控制器95命令读数装置65的活动,并且调节发光二极管(LED)90的光照能量输出,如图5b的左下部所示。
同样,光电二极管92接收光照能量,并且将这种能量转化成传递给前置放大器79的信号,然后传递到微控制器95。最终,将数据输出或将分析结果显示在图5所示的LCD显示器74上。
发光辐射的波长应当选择成,使它处在检测器(光电二极管)具有明显反应的波长范围内(对于硅光电二极管来说,通常为400-1000纳米)。另外,所述光照辐射的波长应当选择成,使它接近在所述横向流动分析中用作标记物的可检测材料的最大吸收波长。
普遍认可的是,在所述分析中被用作标记物或探针的可检测材料,是能通过吸收与可见范围或接近可见范围的光线相互作用的材料。例如,如果所述探针是在浓缩时对裸露的肉眼呈现兰色的物质的话,理想的电磁辐射可能是黄色的。正如本文进一步说明的,颗粒状直接标记物的合适的例子包括金属和金溶胶,非金属元素溶胶(即硒或碳)和彩色乳胶(聚苯乙烯)颗粒。
通过发光二极管90表示的光源可以包括全部的市售部件。合适的例子是市售LED光源,优选选择为可以提供能被检测区42中的浓缩的可检测材料高效吸收的合适波长的光。如果必要的话,可以使用一系列LED光源,它们被轮流提供能量的。
图6表示在图5中示出的本发明的一种实施方案的顶板72的更详细的示意图。如图所示,将具有小块77的膜条76放入第一切口70中。在本发明的某些实施方案中,小块77与第一切口70对齐,以便从膜条76上的参考区83获得读数。然后,一旦获得参考或校正读数,可以将膜条76抬起来,并且改变位置,以便小块77插入第二个切口71。在膜条76上示出了检测区82。然后将检测区82放置在所述孔上(在图6中没有示出该孔),以便获得测试样品的读数。在图6中示出了将膜条76放入其中的通道68。
图7表示沿图6的线7-7的横剖视图。由螺钉67a-b将罩66和顶板72固定在底板78上。在通道68中提供膜条76,以便小块77结合在第一切口70中。在图7中,吸光部件80位于膜条76上面。吸光部件80可以包括用于部件57的所述材料,包括能够吸收光线的几乎任何类型的材料,如黑色或暗色绒屑,毛毡,塑料,金属或其他材料。
本发明的膜式装置包括若干部件,包括膜,样品垫,结合垫和芯吸垫,或以上部件的组合。所述膜通常包括至少两个区,即一个或多个检测区和一个或多个对照或参考区。样品垫接触所述结合垫的一端。
所述分析装置的一种设计包括液体样品流动方向,该方向为样品垫,结合垫,检测区,和垫,通常按从一端到另一端的顺序设置。一般,所述芯吸垫有助于促进通过所述膜条的毛细管抽吸作用和单向液体流动。所述垫将含有分析物的液体沿所述膜从膜的一端到膜的另一端“抽出”。
在本发明中所使用的探针可以包括小球或颗粒。正如本文将要进一步说明的,所述小球或颗粒可以包括乳胶或其他合适材料。在某些用途中,采用普通颗粒,而其他用途可以采用在外表面上具有捕获试剂和/或结合的抗体的颗粒。通常用肉眼可见的或用检测装置可见的染料对所述颗粒进行染色。在其他实施方案中,所述颗粒可以包括吸光材料,如金属溶胶,金,或银颗粒。业已发现,在某些用途中金纳米颗粒是合适的。
在本发明的一种用途中,提供了一种用于进行横向流动分析的系统,用于检测存在于检测液体中的分析物的量。该系统包括采用探针分析物结合复合物,它能够产生可检测信号。另外,提供了一种膜条,并且设计成用于移动含有探针和分析物结合物的检测液体。所述膜条包括检测区,其中,所述检测区具有沉积在它上面的第一种捕获试剂。所述第一种捕获试剂被固定在所述检测区上,并且被设计成用于结合探针分析物结合物,以便固定所述探针分析物结合物,从而在所述检测区内形成夹心复合物。
检测线可以包括固定的第二种捕获试剂(即抗体或其他结合物质),它起着通过结合将未结合的探针固定的作用,以便在捕获线上形成对照探针复合物(即固定物质)。当以这种方式固定了大量探针时,在所述膜条上的一条或多条检测线上出现了可看见的独特的线。可以用预定量的第二种捕获试剂包埋所述对照线。
在某些场合下,在校正的强度水平或对照线(或区),或某些其他参考标准,和所述膜条的检测线之间进行比较,以便计算存在于样品中的分析物的量。该比较步骤是用下面将要进一步说明的读数装置进行的。
在所述分析中所采用的膜条可以是纤维素酯,采用硝酸纤维素通常能提供好的结果,不过,本发明不局限于所述膜条的这种组成。
可以理解的是,可以将本发明设计成检测多种分析物,包括治疗性药物,滥用药物,激素,维生素,葡萄糖蛋白(包括所有类型的抗体),肽,类固醇,细菌或细菌感染,真菌,病毒,寄生虫,细菌的成分或产物,所有类型的过敏原,所有类型的抗原,正常或恶性细胞的产物或成分等。
以下分析物是可以用本发明方法检测的分析物的例子:T.sub.4,T.sub.3,洋地黄毒苷,hCG,胰岛素,茶碱,黄体生成素,导致各种疾病状态或与各种疾病状态相关的生物,如酿脓链球菌(A型),I和II型单纯疱疹病毒,细胞肥大病毒,衣原体,和本领域已知的其他生物。
美国专利号4366241(Tom等)在第19-26栏列举了多种所关心的潜在分析物,它们是免疫学对的成员,包括蛋白,凝血因子,激素,微生物,药用制剂,和维生素。所述分析物中的任意一种都适合用作本发明的分析物。
可以检测的优选配体或分析物的其他例子包括以下成分:人骨骼碱性磷酸酶抗原(HBAPAg);人绒毛膜促性腺素(hCG);人黄体生长素(hLH);人促滤泡激素(hFSH);肌酸磷酸激酶MB同工酶;铁蛋白;癌胚抗原(CEA);前列腺特异抗原(PSA);CA-549(乳腺癌抗原);乙型肝炎表面抗原(HBsAg);乙型肝炎表面抗体(HBsAb);乙型肝炎核心抗原(HBcAg);乙型肝炎核心抗体(HBcAb);甲型肝炎病毒抗体;人类免疫缺陷病毒抗原HIV I,如gp120,p66,p41,p31,p24或p17;HIV II的p41抗原;和针对上述配体中任意一种的相应的抗配体(优选单克隆抗体)。在以下文献中对HIV抗原进行了更全面的说明:美国专利号5120662,和Gelderblood等人的Virology156:171-176,1987,这两份文件都被收作本文参考。
在本文中,术语“探针”一般表示能够在横向流动分析中将分析物携带到检测部位或检测区的结构,它可以是或不是颗粒或微颗粒形式的。另外,在本文中,术语“探针-结合物”表示在横向流动分析中能够携带分析物以便形成探针-结合物复合物的物质,它能在膜条的检测区中固定第一种捕获试剂,以便在所述检测区形成“夹心复合物”。
在本文中,术语“微颗粒”更具体的讲是表示一种特定类型的探针,并且可以包括任何珠子或探针,抗体可以与它们结合,这种结合可以是共价的或非共价的,如通过吸附结合。用于定量分析中的某些颗粒的其他要求是,所述颗粒能产生一种信号,通常是光吸收,这会导致所述颗粒所在的区域具有不同于所述膜的其他部位的信号。
可选择地,将金属颗粒或金属用作本发明的探针。所述颗粒是以具有大体上一致的直径的微球体形式通过商业渠道获得的,如从诸如英国Cardiff,British Biocell International的公司购买。
本文所使用的短语“膜”或“膜条”表示采用膜和一种或多种试剂检测测试溶液中优选是含水的测试溶液中所关心的分析物的浓度的检测装置或条。与所述膜式装置结合的至少一种试剂是所关心的分析物的结合配偶体。
用于本发明的乳胶微颗粒是以具有大体上一致的直径的聚合微球体的形式(以下称之为“聚合微球体”)通过商业渠道获得的,如从印地安纳州的Bangs Laboratories of Carmel或米歇根州的Midland的Dow Chemical公司购买。尽管任何能够吸附或与结合配偶体共价结合的聚合微球体都可用于本发明,但所述聚合微球体通常是由下列一组的一个或多个成员组成:聚苯乙烯,丁二烯苯乙烯,苯乙烯丙烯酸-乙烯基三元共聚物,聚甲基丙烯酸甲酯,聚乙基丙烯酸乙酯,苯乙烯-马来酸酐共聚物,聚乙酸乙烯酯,聚乙烯基吡啶,聚二乙烯基苯,聚对苯二甲酸丁二醇酯,丙烯腈,氯乙烯-丙烯酸等或它们的醛,羧基,氨基,羟基,或酰肼衍生物。
诸如聚苯乙烯的非衍生的聚合微球体是疏水性的,并且能被动吸收其他疏水性分子,包括大部分蛋白和抗体。在以下Cantarero等人的文献中提供了用于将蛋白或多肽吸附在疏水性颗粒上的技术:“苯乙烯对蛋白的吸附特征及其在固相免疫分析中的作用”,AnalyticalBiochemistry 105,375-382,(1980);和Bangs,“乳胶免疫分析”,J.Clin.Immunoassay,13,127-131(1980),这两份文献都被收作本文参考。
一般来说,在以下文献中还披露了用于将分子吸附在聚合微球体上的各种方法:在Bangs,L.B.中,“均匀乳胶颗粒”,在第41届美国临床化学协会的国家会议专题研讨会上提供,1989,并且可以以印刷品形式从Seragen Diagnostics Inc.获得,Indianapolis Ind.;或Galloway,R.J.“微颗粒检测和免疫分析的发展”,即印地安纳的Seradyn Inc.,这些文献被收作本文参考。
所述检测溶液可以是生物学流体的成分,如从植物或动物,优选哺乳动物,更优选人体内提取的,稀释的,或浓缩的成分。特别优选的生物学流体是血清,血浆,尿液,腹水,腹膜液,羊膜液,滑液,脑脊液等,或它们的浓缩物或稀释液。
在实施本发明时,校正和样品检测可以在同一时间在大体上相同的条件下进行,因此,提供了高度可靠的定量结果,和较高的灵敏度。
本发明还可用于半定量检测。作为多个控制线,提供了一定范围的信号强度,可以将所述检测线的信号强度(即例如可见的),与对照线进行比较。根据所述检测线所处在的强度范围,可以确定所述分析物的可能的浓度范围。所述探针可以是用任何信号发生物质标记过的乳胶珠,或所述标记过的乳胶珠进一步与抗体结合。
本领域普通技术人员可以理解的是,本讨论仅仅是对典型实施方案的说明,并且,并不是要限定本发明的较宽的范围,所述较宽的范围体现在所述典型结构中,本发明是通过所附权利要求书的形式呈现的。
Claims (15)
1.一种改进的横向流动分析读数装置,该读数装置被设计成能够从横向流动分析膜条中检测分析结果,所述分析结果是通过检测分析物在所述膜条的检测区内的结合显现的,该装置包括:
(a)具有外部和内部的外壳;
(b)所述外壳内的容纳口,用于将膜条直接从所述外壳的外部纳入所述读数装置外壳的内部,所述容纳口具有一个遮光结构;
(c)读数机构,该机构包括:
(i)电磁辐射源;
(ii)一个或多个能够检测电磁辐射强度的传感器,其中,电磁辐射源和传感器位于所述读数机构内,以便当所述膜条被纳入所述容纳口时,并且由所述遮光结构封闭时,所述电磁辐射首先照射在所述膜条上的检测区,然后再照射在所述传感器上,其中该电磁辐射在进入容纳口之前通过一孔,该孔的尺寸大致为检测区的尺寸。
2.如权利要求1的读数装置,其中,所述容纳口包括压在所述膜条上的压力板。
3.如权利要求2的读数装置,其中,所述压力板是弹簧加载的。
4.如权利要求1-3任一项的读数装置,其中,在所述容纳口内提供吸光部件,以便吸收散射光线。
5.如权利要求4的读数装置,其中,所述吸光部件包括位于容纳口内的膜条附近的吸收垫。
6.如权利要求5的读数装置,其中,所述吸光部件包括柔性的和顺应性的毛毡材料。
7.如权利要求5的读数装置,其中,所述电磁辐射首先通过孔,然后再进入所述容纳口。
8.如权利要求1-3中任一项的读数装置,其中,所述孔是长形的。
9.如权利要求1-3中任一项的读数装置,其中,所述孔是圆形的。
10.如权利要求1-3中任一项的读数装置,其中,所述容纳口包括用于参考读数的第一个停止位置,和用于样品读数的第二个停止位置。
11.一种检测盒,包括权利要求1-10中任一项的横向流动分析读数装置,以及一个分析装置,该分析装置包括多孔液体可渗透的膜条,其中,所述多孔液体可渗透膜条的厚度是电磁辐射能够穿透的,其中,所述分析结果是通过可检测分析物与固定在所述检测区上的结合试剂的直接或间接地特殊结合显现的。
12.如权利要求11的检测盒,其中,所述读数机构部分地通过用传感器检测从以下区域反射的电磁辐射来确定所述分析结果:
a)所述膜条的检测区;和
b)所述膜条上的校正区,
其中,对在所述膜条的检测区和所述膜条的校正区中测定的值进行比较。
13.一种用于进行横向流动分析,以便检测存在于检测液体中分析物的量的系统,该系统包括:
(a)能够产生可检测信号的探针-分析物结合物;
(b)膜条,所述膜条包括检测区,所述检测区具有沉积在它上面的第一种捕获试剂,其中,所述第一种捕获试剂被设计成用于结合探针-分析物结合物,以便固定所述探针-分析物结合物,从而在所述检测区内形成夹心复合物;和
(c)根据权利要求1-10中任一项的读数装置。
14.一种用于进行横向流动分析以便检测在检测液体中分析物的数量的方法,该方法包括:
(a)在膜条上提供探针-结合物,将所述探针-结合物设计成能产生可检测信号;
(b)在所述膜条上提供分析物;
(c)将所述探针-结合物结合在所述分析物上,以便形成探针-分析物结合物复合物;
(d)其中,将所述膜条设计成将含有探针和探针-分析物结合物复合物的检测液体固定,所述膜条包括检测区;
(e)在所述检测区提供第一种捕获试剂,其中,所述第一种捕获试剂是固定的,并且被设计成能结合所述探针-分析物结合物复合物,以便通过在所述检测区内形成夹心复合物,固定所述探针-分析物结合物复合物;
(f)提供读数装置,将所述读数装置设计成检测来自所述膜条的分析结果,所述读数装置包括具有外部和内部,以及具有遮光结构的容纳口的外壳,所述容纳口结合在所述外壳内,用于将所述膜直接从读数装置外壳外部纳入读数装置外壳内部;
(g)阻止环境光线到达所述膜检测区,以便提高灵敏度;
(h)其中,所述电磁辐射源和传感器是这样定位的,使得当所述膜条被纳入所述容纳口时,电磁辐射首先照射在所述膜条上的检测区,然后到所述传感器;
(i)设置使电磁辐射在进入容纳口之前通过一孔,所述孔的尺寸大致为检测区的尺寸。
15.根据权利要求14所述的方法,其特征在于,还包括在容纳口中设置的一吸光部件以吸收散射光,该吸光部件包括吸收垫,该吸收垫靠近在容纳口内的膜条并且在使用中位于膜条的上表面上,该吸收垫完全覆盖这样的区域:在该区域下面该膜条被来自该膜条下面的光照射。
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2001
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN105408749A (zh) * | 2013-08-02 | 2016-03-16 | 电化生研株式会社 | 免疫色谱法中使用光反射材料的检测光的增强方法 |
Also Published As
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EP1459054A1 (en) | 2004-09-22 |
BR0214478A (pt) | 2004-12-28 |
KR20040071750A (ko) | 2004-08-12 |
DE60223402T2 (de) | 2008-08-28 |
ES2291530T3 (es) | 2008-03-01 |
WO2003058220A1 (en) | 2003-07-17 |
US20030119202A1 (en) | 2003-06-26 |
US8367013B2 (en) | 2013-02-05 |
DE60223402D1 (de) | 2007-12-20 |
ATE377752T1 (de) | 2007-11-15 |
TW200305011A (en) | 2003-10-16 |
CN1608201A (zh) | 2005-04-20 |
CA2471370A1 (en) | 2003-07-17 |
TWI220924B (en) | 2004-09-11 |
EP1459054B1 (en) | 2007-11-07 |
MXPA04006171A (es) | 2004-12-06 |
AU2002363945A1 (en) | 2003-07-24 |
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