CN1146557A - 定量检测免疫层析条上的分析物 - Google Patents
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Abstract
本发明公开了一种改进的利用免疫层析技术测定试样中分析物浓度的方法,包括借助仪器装置(例如反射分光光度计)定量测定来自捕获分析物标记结合对复合物的信号。在一个优选的实施例中,测定捕获复合物和在免疫层析条的单独区域中被捕获的未复合标记结合对的反射度读数,并利用这两个反射度的比率对分析方法进行定量。
Description
免疫层析条形式越来越广泛地用于采用目视检测方案的定量和半定量分析。这种类型的免疫分析包括将怀疑含有待测分析物的液体试样施加到免疫层析试条的点样区。该试条由一种基质材料构成,试液和悬浮或溶解于其中的分析物通过毛细作用可经过这种基质材料,从点样区流到检测区,而在检测区是否出现目测信号则表明分析物的存在与否。特别是,这种试条还包括使待测分析物与它的带有可检测的标记物的特异性结合对发生免疫特异性结合的试剂。在这种设计中,正如美国专利4,446,232中所公开的那样,试条含有一种被标记到可与分析物作用的流动结合对上的酶,此结合对位于点样区的下游区域中。如果试样中存在分析物,那么它将与标记结合对结合,形成一个复合物,该复合物沿试条流到含有酶标记物的底物的检测区,这种底物在酶标记物的存在下能提供一个颜色反应。试条含有一个可固定分析物的区域,从而使标记结合对在样品中没有分析物存在时不与分析物结合而被捕获,借此避免到达检测区。目前已经公开了这种技术的各种改进形式,所以这些改进形式都包括一些竞争的特异性结合系统,在此系统中通过检测检测区域中是否有标记结合对来测定试样中是否存在分析物。在美国专利4,868,108中公开了一种类似的方案:在检测区域加入一种酶标记结合对的固定化捕获剂来浓缩酶标记物,从而使它与酶底物的反应能力加强,借此使分析更加灵敏。
并不是所有的免疫层析方案都依靠酶标记的结合对/酶底物来提供分析物的检测信号。在美国专利4,806,311中公开了一种用于对分析物进行特异性结合分析测定的多区域试验装置、一种固定化结合对以及一个接收从试剂区迁移过来的标记试剂的检测区域。检测区含有这种标记试剂的结合物的固定形式。此标记试剂带有一个可检测的化学基团,该化学基团具有可检测的物理性质,根据其自身的物理性质即可检测而无需和另一物质发生化学反应。这些基团的例子包括彩色荧光剂、磷光分子、放射性同位素和电活化部分。
美国专利4,313,734中描述了利用金溶胶作为检测抗体的标记物,无需化学变化即可检测。
免疫层析条形式提供了一种用于测定各种分析物(无论是抗原还是抗体)的可视系统但却具有局限性,即,它们最多只能产生半定量结果,而对于一些分析物却需要定量结果。因此,本发明的一个目的是提供一种利用免疫层析条形式对分析物进行定量测定的方法。
本发明涉及一种改进的用于测定试液中分析物的方法,包括将试液施加到免疫层析基质上,试液和分析物(如果存在的话)通过毛细作用流经基质并且基质中含有一个可与分析物作用的标记结合对。基质,通常在试条的形式中,还含有至少一个检测区,在该区域中通过利用特异性结合对检测标记物来测定是否存在分析物。这种改进在于可利用一种装置测定标记物的浓度,此装置具有一个能测定检测区域中标记物浓度的检测器。
在本发明的一个优选实施例中,提供了一种试条,该试条包括一个具有一个第一区域和至少一个第二区域的条形物,其中第一区域含有可与分析物作用、带有可检测的标记物并能与分析物反应形成分析物/标记结合对复合物的流动特异性结合对,第二区域含有一种固定化分析物或其类似物。此处所用的“类似物”一词意指任何能通过特异性结合对的活性部位进行结合的物质。
上述的试条可如下展开:将怀疑含有分析物的试液样品施加到其上从而使样品与流动的、可与分析物作用的标记特异性结合对接触,借此流体试样中存在的分析物与标记特异性结合对结合而形成复合物;剩下多余的未反应的标记结合对不再进行反应,从而流体试样通过毛细作用将分析物/标记结合对共轭物以及未反应的标记结合对沿试条运载到含有固定化分析物或其类似物的第二区域,在此区域未反应的标记结合对与固定化分析物结合,其中固定化分析物与流体试样中的分析物的浓度成反比。
展开试条在具有能够测定来自可检测标记物的信号的检测器的装置上读数,以便测定来自第二区域中的标记结合对的信号。通过将来自可检测标记物的信号与以类似方法利用含有已知浓度分析物的流体试样进行的测定比较来测定流体试样中分析物的浓度。
通过使试条配有一个第三区域可增强该测定方法的灵敏度,此第三区域含有用于固定由分析物和标记结合对形成的复合物的试剂。例如,如果利用标记的小鼠抗体(IgG)中的标记结合对,那么这种小鼠抗体与分析物的复合物在固定有山羊抗-鼠IgG的区域被捕获。通过测定固定在这个第三区域中的可检测标记物的信号以及第二区域和第三区域中被标记结合物的信号比,可校正由被标记共轭物的不匀沉积和/或穿过基质的非均匀流体流动产生的偏差。
可如下实施本发明:首先提供一个试验基质,流体试样可通过毛细作用流经该基质。特别是,基质将制成试条的形式,试液可水平流经该基质,即使基质分层设置,试液也可从顶部到底部或从底部到顶部垂直流经该基质。以下将集中讨论试条形式。
试条可用试液和其中含有的分析物能通过毛细作用流经的任何基质材料制成。基质是一种能够非吸水性横向流动的材料。美国专利4,943,522中描述了这种作为液流的流动,其中液体的所有溶解或分散的组分以基本相同的速率和相对不减弱的流动穿过基质,这与一种或多种组分的优先保留的情况相反,如果基质材料能吸附或吸收一种或多种组分,就会发生后一种情况。这种基质材料的一个例子是由Porex Technologies of Fairburn,GA生产的高密度或超高分子量聚乙烯片材。同样适于用作制成层析条的基质材料是诸如纸、硝基纤维素和尼龙这些吸水性材料。
各种免疫层析条形式都适于同本发明一起使用。美国专利4,446,232中公开了种特别适合的形式,在该专利中描述了一种用于测定抗原存在的装置,该装置包括一个具有一个第一区域的基质材料条,在第一区域中具有固定化分析物和特异于待测分析物的酶联抗体。当标记抗体与渗入第一区域的分析物反应时就可流入第二区域,但是当试液中无分析物存在时,标记抗体由于同固定化分析物作用而结合在第一区域不再流动。虽然这种形式的试条可以被设计成用于抗体作为分析物来检测其存在的,但是典型的分析物是抗原。美国专利4,868,108中公开了这种形式的变型。在另一变型中,酶底物沉积在第二个含有固定化抗体的区域中,借此捕获酶标记的抗体与分析物形成的复合物。虽然由于本发明并不局限于酶与其底物的相互作用来提供可检测的信号而可采用任何物理上可检测的信号,但是这种形式特别适用于本发明。这样,通过将共轭物固定在一个离散区域中(该区域位于试条上结合有可与分析物作用的标记结合对的区域下游),可提供两个区域,从这两个区域中能测定到可检测标记物的物理上可检测的性质以便测定标记物的浓度。通过测定来自第二区域(含有作为捕获得剂的固定化分析物)中的可检测标记物的物理上可检测性质的信号以及来自第三区域(在该区域,针对标记结合对的固定化抗体是捕获剂)中的标记物的物理上可检测性质的信号并测定这两个信号的比率,可增加测定分析物浓度的试验的准确性。准确性之所以增加是因为这种技术校正了由标记共轭物的沉积和/或不均匀流体流经基质所产生的偏差。更具体地说,由于上述标记共轭物的沉积和不均匀流体流动的偏差通常较小但却相当重要,所以它们基本上不干扰结合平衡。因此,这两个结合区域中的信号比比任何一个区域中本身的信号更能准确地测定分析物的浓度。
在本发明的一个优选实施例中,提供了一种反射分光光度计,该光度计带有使试条或检测器彼此相对移动的装置例如样品桌,试条放在样品桌上能横向移动到检测器读数头之下的位置上。在可检测的物理性质是预定波长处的光的反射度的情况下,检测器是分光光度计。这种技术有助于准确定量没有精确位于分光光度计的检测装置的相关位置上的试条区域。更具体地说,试条相对于检测器的位置可以由微处理器控制,从而可测定任向所需区域的反射度。
下面将借助几个例子更充分地描述实施本发明的方法。例1用单个封闭带的形式对HSA进行定量测定
制备一免疫层析条,该试条在-ImmunodyneTM尼龙膜上含有一固定化HSA封闭带和大面积抗-HSA:金溶胶共轭物。此试条如图1所示,其中条10包括封闭带3以及位于封闭带3之前的包含抗-HSA:金溶胶的区域5和点样区7。如下制备这些试条:
将-4.2×12.6厘米的ImmunodyneR膜片放在Comag薄层色谱(ILC)剥离装置上,并且其长边与机座平行,从Y轴的0位置处向上偏置1cm。然后,在磷酸盐缓冲盐水(PBS;0.137M氯化钠,0.0027M氯化钾,0.010M磷酸钾,PH7.4)中制备液度为10mg/mL的人血清白蛋白(HSA)溶液。如下设置TLC剥离器,在3.5cm的Y位置处剥离-6cm长的10mg/mLHSA溶液带:
(a)板=90,(b)带=60,(c)秒/μL+6,
(d)体积=6μL由此得到一6cm长、大约1mm宽的带。因此剥离密度是10μL/cm2而HSA的密度为100μg/cm2。
3分钟以后将膜从TLC剥离器上拿开,放入一盛有0.5%酪蛋白(Hammerstein from Schlesinger)磷酸盐缓冲盐水(pH7.4from Sigma)的平塑料盘中并在一轨道摇动器中轻轻摇动30分钟。
此时制备一含有0.02%叠氮化钠、0.02%吐温20及0.1%PEG20的PBS洗涤缓冲剂。然后将膜从酪蛋白封闭液中拿出,用25mL洗涤缓冲剂洗涤两次,总共洗涤30分钟并在轨道缓冲剂中轻轻摇动,然后将膜从洗涤缓冲剂中拿出并在室温下干燥过夜。
将2.0mL 10mg/mL的酸性氯化金-水化物(HCl4Au·H2O)加入到柠檬酸酸钠(0.00155M)的100℃回流溶液中来制备金溶胶。继续回流30分钟,然后冷却并用0.2μM的滤纸过滤。将240μg的针对人血清白蛋白的单克隆抗体和50μL0.1M的碳酸钾加入到10mL上述制备的金溶胶溶液中来制备抗体-金溶胶共轭物(Ab:gold sol),并将混合物剧烈搅拌15分钟,然后加入0.5mL 1%(w/v)的PEG-20,再剧烈搅拌10分钟。此时,加入1.0mL 10%的牛血清白蛋白(BSA)水溶液并将混合物剧烈搅拌10分钟。通过在20℃ 14,500Xg下离心30分钟分离出抗体:金溶胶,然后使其悬浮在洗涤缓冲剂(1%BSA,0.05%PEG-20,2mM硼酸钠,pH=9.0)中洗涤10次并如上所述进行离心分离。最后离心之后,将抗体:金溶胶悬浮在1.0mL的洗涤缓冲剂中并在4℃下保存。
再将上述的干燥膜放在TLC剥离器上,如上所述在Y方向上偏置1cm。制备40μL抗体:金溶胶、20μL 4%酪蛋白及20μL1%Methocel(k4M)+0.6%聚乙烯醇(PVA)的混合物并且如上所述在2.3和2.9cm的Y位置之间剥离7条相邻带。使试条在室温下干燥并在使用之前切成0.5cm宽的窄条。
用一超滤膜过滤中等比重(S.G.+1.017)的尿池,这种滤膜能阻挡大于30,000道尔顿的蛋白质。通过将滤液与已知浓度的HSA溶液掺合可制备各种已知浓度的HSA溶液。
可如下展开试条:将试条垂直悬在掺合有尿滤液的HSA溶液中深度约达0.5cm(在试条含有抗体:金溶胶共轭带的那一端)并保持5-10分钟以使液体到达试条的顶部。在室温下空气干燥这些试条,然后将其置于塑料trycite手柄上,并进行分析。
利用含有0,1,2,3和5mg/dL HSA的中等比重尿超滤液样品展开试条。在每两次测量之间都将试条端部切去1毫米,用CT100反射分光计扫描经过样模拟的试条,通过测定557nm处的反射度来读取用每个样品浓度展开的试条。
更具体地说,在将试条用样品液体展开并在室温下进行空气干燥之后,利用双边粘合剂将其粘在塑料手柄上。从由于结合有抗体:金溶胶而可目测的HSA带3(图1)将塑料/膜层的叠层修剪成7mm(朝向施加样品的那一端)。然后将试条放在CT100反射分光计的读数台上并将试条推到顶站。在此位置10 th 衬垫位置的读数区域是从试条那一端起2.5mm。然后从试条一端切下1mm并将试条推到读数台的端部,再测定10 th 衬垫位置的反射度。再读取10 th 衬垫位置并重复此过程直到塑料和膜的叠层的一端与经过HSA带3mm的点对应。由于没有软件可完成此任务,可利用此技术使读数区(HSA带)相对于读数头(检测器)移动。利用适当的软件,使读数台和塑料以及膜的叠层经过读数头移动,可扫描试条的反射度。
本实验的结果以图表的方式示于附图2中。
从图2中可测得用含有不同浓度HSA的尿样展开的免疫层析条的反射度扫描谷的深度与HSA的浓度成正比并且从反射度可看出HSA的剂量反应。即使金溶胶带没有覆盖整个读数区,当此带在读数区时反射度也降低10-15%。尽管许多高反射(白)区可随金溶胶带一起测定,但是10-15%的反射度变化也能检测出。
如果将带有狭缝的屏蔽放到读数头区域中,那么由于高反射白区不能与金溶胶带同时处于读数区中所以反射度范围将显著增大。此增大的反射度可更好地区别分析物(HSA)的浓度。利用步进电机,可使分光光度计的试条台慢慢通过试条上的任何区域,同时由于步进电机一次只能移动每一次循环的一小部分,因此可给出连续的读数,从而为测定谷反射度或谷中的各个区域提供的良好的分辨率。例II用含有封闭带和捕获区域的形式对HSA进行定量测定
按照例I的方法和图3的形式制备一条免疫层析条。参考图3,试条10具有固定化HSA封闭带3、小鼠抗-HSA:金溶胶共轭物区域5和固定化山羊抗-小鼠IgG抗的捕获带9。在制备此带时,在0.135M氯化钠中制备浓度为5mg/mL的山羊抗-小鼠IgG(sigma 8770)溶液。如上所述在4.0cm的Y距离处进行剥离。剥离密度为50μg IgG/cm2。当将点样区7浸到含有HSA的样品中的深度小于需浸渍共轭物区域5的深度时,流体将通过毛细作用从样品向上流动。样品中的HSA将与共轭物区域中的金溶胶:抗HSA复合并将随共轭物一起向试条顶部移动,这些共轭物是游离的,因为共轭物相对样品中的HSA是过量的,它找不到任何HSA可供结合。游离的共轭物将结合封闭带3中的固定化HSA同时金溶胶-抗HSA:HSA复合物继续向试条顶部流动,在试条上它将与捕获带9中的固定化山羊抗小鼠IgG抗体结合。
用含有0,0.5,0.8,1.0和2.0mg/dL HSA的中等SG尿超滤液展开这种形式的试条。用每个HSA浓度重复展开试条。利用将这些带目测定位在10 th 衬垫位置中心的方法在CT100上采集557nm处的反射度。10 th 衬垫位置占据邻近MultistixR10SG尿试条制品一端的5mm的部分。Multistix 10SG制品是一大约10.9cm长、5mm宽、0.5mm厚的塑料片。10个含有干试剂的纸垫(每个5mm×5mm)可以附着在其上。10 th 垫平坦地与塑料片的一端对齐,并且垫与垫之间的间隔为2.5mm,而在没有垫并用作把手部分的另一端剩余3.4cm的塑料片。在图4a中,以反射度为单位画出了本试验的结果。在此图中,反射度来自固定化HSA带3(图3),对HSA的剂量反应以反射度(R)为单位与两个偏离值成线性。正如图4b所示的,山羊抗-小鼠IgG带9(图3)对HSA的反应更分散并且大多数反射度点出现在0-0.5mg/mL HSA之间。然而,当HSA带的反射度值与山羊抗-小鼠IgG带的反射度值成比率时,正如从图4c所测得的,变异性减小,图4C是HSA浓度相对于HSA带的反射度与山羊抗-小鼠IgG带的反射度的比率的曲线。在图4C中可观察到对HSA浓度的弯曲但却光滑的剂量反应。这个比率可以倒过来,这相当于取每个比值的倒数,这样做与比率本身具有相同的作用。这样,测定这两个反射度值的比率可用于校正试剂制备期间共轭物沉积的差异以及在用样品展开试条期间流体流动中的任何不均匀性,即,如果一个试条上的金溶胶:抗HSA比另一试条上的少,那么这两条带的比率将提供一个结果用于校正制备中的不均匀性。用类似的方法可校正试条展开期间流体流动中的不均匀性。例III用双封闭带形式对HSA和IgG进行定量测定
制备能测定两种物质即人血清白蛋白(HSA)和人(H)IgG的免疫层析条以便对每个分析物各自进行定量。按照图5的设计用ImmunodyneTM尼龙制备此试条,使其包括点样区1,其次是含有金溶胶标记的抗-HSA和金溶胶标记的抗-(H)IgG共轭物的区域3。该试条包括两个封闭带,即含有固定化HSA的第一封闭带5和含有固定化(H)IgG的第2封闭带7。
用含有0,5,10,15,20,30和40mg/mL HSA以及0 IgG或100mg/mL(H)IgG的中等SG尿的超滤液样品展开试条。通过物理切割试条并使试条对准MultistixRSG试条的10 th 衬垫位置,在CT100反射分光计上测定557nm处的反射度,得到各个反射度读数以此来检查用每个HSA浓度展开的两个试条。图6用图表形式示出了含有HSA和(H)IgG封闭带的样品的反射度,所用试样不含IgG。图7示出了含有100mg/L(H)IgG和不同浓度HSA的试样的类似数据。从图6的数据可确定HSA封闭带的反射度与样品中的HSA浓度成正比并且由于样品中无(H)IgG使得金溶胶:抗(H)IgG结合到(H)IgG封闭带上而使(H)IgG封闭带的反射度大约为0.88(基于最大总反射度为1.0)。图7中的数据如图6中的数据那样表明反射度与HSA浓度成正比,但是由于样品中含有100mg/L的(H)IgG使得(H)IgG的反射度稍高(0.91-0.92)。(H)IgG结合到金溶胶-抗(H)IgG共轭物上而使共轭物不再与(H)IgG封闭带中的固定化(H)IgG结合。这表明无论(H)IgG存在与否对HSA的剂量反应都是相同的,并且有一个独立的对(H)IgG的剂量反应。因此,通过利用一个带有针对两种不同分析物的混合金溶胶抗体共轭物的试条,在具有这些分析物或类似物的固定化带的隔离区域中,可得到独立的用仪器可检测的对每种分析物的剂量反应。这很重要,因为如此可利用一个免疫层析条定量多种分析物。
Claims (17)
1.一种测定流体试样中分析物的方法,包括将流体施加到一种免疫层析基质上,使流体试样通过毛细作用沿基质向上流动,该基质含有一种可与分析物作用的标记结合对和至少一个检测区域,在该检测区域通过利用标记结合对检测所含的标记物来测定分析物,其特征在于此方法包括利用一种带有检测器的仪器测定来自标记物的信号,这种检测器能够测定检测区域中标记物的信号。
2.一种用于测定流体试样中一种或多种分析物浓度的方法,包括:
a)提供一种包括一种基质的试条,流体试样通过毛细作用可流经该基质,其中试条的第一区域含有可与分析物作用的流动特异性结合对,这种结合对带有可检测的标记物并能与分析物反应形成分析物/标记结合对复合物,试条还具有至少一个含有固定化分析物或其类似物的第二区域。
b)按如下步骤展开试条:将怀疑含有分析物的流体试样施加到试条上,由此使得分析物与可同其作用的流动的标记特异性结合对接触,从而流体试样中存在的分析物与标记特异性结合对结合形成复合物,剩余过量的未反应的标记结合对不再进行反应,流体试样通过毛细作用将分析物/标记结合对复合物以及未反应的标记结合对沿试条运载到含有固定化分析物或其类似物的第二区域,在此区域未反应的标记结合对与固定化分析物结合,其中固定化分析物与流体试样中的分析物的浓度成反比,
c)在一种带有检测器的仪器上读取被展开的试条,该检测器能够测定来自可检测的标记物的信号以便测定第二区域中的标记结合对的浓度,以及
d)通过将步骤C)测定的来自可检测标记物的信号与以类似方法利用含有已知浓度分析物的流体样品所得到的信号测定值进行比较来测定流体试样中分析物的浓度。
3.如权利要求2所述的方法,其特征在于试条还有一个第三区域,该第三区域含有用于固定分析物和标记结合对形成的复合物并测定来自固定在这个第三区域中的可检测标记物的信号的试剂,由此可测定来自固定在第二区域中的标记结合对和第三区域中的固定化复合物的信号比。
4.如权利要求2所述的方法,其特征在于所述仪器包括用于使试条或检测器彼此相对移动的装置,以便准确地定量没有精确地位于仪器的相关位置上的区域。
5.如权利要求2所述的方法,其特征在于在试条的相同或不同区域中有两个或多个流动的标记结合对和两个或多个含有固定化分析物或其类似物的区域,从而在一个试验中可测定两个或多个分析物。
6.如权利要求2所述的方法,其特征在于所述仪器是一种反射分光光度计。
7.如权利要求2所述的方法,其特征在于基质是由一种能非吸水性横向流动的材料或一种吸水性材料构成。
8.如权利要求2所述的方法,其特征在于所述分析物是人血清白蛋白(HSA)。
9.如权利要求2所述的方法,其特征在于所述可检测的标记物是一种有色物质。
10.如权利要求9所述的方法,其特征在于所述有色物质是一种金溶胶。
11.如权利要求2所述的方法,其特征在于所述被标记结合对是一种抗体。
12.一种用于定量测定流体试样中一个或多个分析物浓度的方法,包括以下步骤:
a)提供一种包括一种基质的试条,流体试样通过毛细作用可以流经该基质,所述试条具有一个含有可与分析物质作用的流动特异性结合对的第一区域,所述结合对带有可检测的标记物并能与分析物反应形成分析物/标记结合对复合物,所述试条还具有至少一个含有固定化分析物或其类似物的第二区域和至少一个含有用于固定分析物和标记结合对形成的复合物的第三区域;
b)可按如下方式展开试条:将怀疑含有分析物的流体试样施加到试条上,由此使得分析物与流动的特异性结合对接触,从而流体试样中存在的分析物与标记特异性结合对结合形成复合物,剩余过量的未反应的标记结合对不再进行反应,流体试样通过毛细作用将分析物/标记结合对复合物和未反应的标记结合对沿试条运载到含有固定化分析物或其类似物的第二区域,在此区域未反应的标记结合对与固定化分析物结合,其中固定化分析物与流体试样中分析物的浓度成反比,通过毛细作用将分析物/标记结合对复合物运载到第三区域,复合物在此区域被固定剂捕获;
c)在一种带有检测器的仪器上读取被展开试条的第二区域,该检测器能够测定来自可检测的标记物的信号以便测定第二区域中标记结合对的浓度并以类似的方式读取被展开试条的第三区域以便测定来自该试条的第三区域中标记结合对的信号;
d)计算来自固定在第二区域中的标记结合对的信号同来自第三区域中的标记结合对的信号的比率;以及
e)通过将步骤c中测定的信号比与以类似方法用含有已知浓度分析物的流体样品所测得的信号比进行比较来确定流体试样中分析物的浓度。
13.如权利要求12所述的方法,其特征在于所述测定仪器是一种反射分光光度计。
14.如权利要求12所述的方法,其特征在于所述基质是由一种能够非吸水性横向流动的材料或一种吸水性材料构成。
15.如权利要求12所述的方法,其特征在于所述分析物是人血清白蛋白(HSA)。
16.如权利要求12所述的方法,其特征在于所述可检测的标记物是金溶胶。
17.如权利要求12所述的方法,其特征在于所述标记结合对是一种抗体。
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US08/380,119 US5569608A (en) | 1995-01-30 | 1995-01-30 | Quantitative detection of analytes on immunochromatographic strips |
US380,119 | 1995-01-30 |
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CN96104311A Pending CN1146557A (zh) | 1995-01-30 | 1996-01-29 | 定量检测免疫层析条上的分析物 |
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EP (1) | EP0724157A2 (zh) |
JP (1) | JPH08240591A (zh) |
KR (1) | KR960029790A (zh) |
CN (1) | CN1146557A (zh) |
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-
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- 1996-01-10 CA CA002166913A patent/CA2166913A1/en not_active Abandoned
- 1996-01-17 ZA ZA96357A patent/ZA96357B/xx unknown
- 1996-01-17 EP EP96100599A patent/EP0724157A2/en not_active Withdrawn
- 1996-01-27 KR KR1019960001777A patent/KR960029790A/ko not_active Application Discontinuation
- 1996-01-29 JP JP8013273A patent/JPH08240591A/ja active Pending
- 1996-01-29 AU AU42226/96A patent/AU4222696A/en not_active Abandoned
- 1996-01-29 CN CN96104311A patent/CN1146557A/zh active Pending
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CN100347547C (zh) * | 1999-05-13 | 2007-11-07 | 松下电器产业株式会社 | 色谱定量测定装置、色谱定量测定方法及其使用的色谱试验片 |
CN100430726C (zh) * | 1999-10-25 | 2008-11-05 | 松下电器产业株式会社 | 免疫色谱试验片及色谱分析方法 |
CN100533147C (zh) * | 2000-05-26 | 2009-08-26 | 松下电器产业株式会社 | 生物传感器 |
US7678566B2 (en) | 2000-09-25 | 2010-03-16 | Panasonic Corporation | Device for chromatographic quantitative measurement |
US8822230B2 (en) | 2000-09-25 | 2014-09-02 | Panasonic Healthcare Co., Ltd. | Chromatography quantitative measuring apparatus |
US8778698B2 (en) | 2000-09-25 | 2014-07-15 | Panasonic Healthcare Co., Ltd. | Chromatography quantitative measuring apparatus |
US8722424B2 (en) | 2000-09-25 | 2014-05-13 | Panasonic Corporation | Chromatography quantitative measuring apparatus |
US8722425B2 (en) | 2000-09-25 | 2014-05-13 | Panasonic Corporation | Chromatography quantitative measuring apparatus |
CN100451652C (zh) * | 2003-03-28 | 2009-01-14 | 安尼生物科技公司 | 多通道测试装置、其生产方法及其用途 |
CN1954214B (zh) * | 2004-03-30 | 2012-07-04 | 通用电气医疗集团生物科学公司 | 侧流格式、材料和方法 |
CN1954212B (zh) * | 2004-05-12 | 2013-03-13 | 霍夫曼-拉罗奇有限公司 | 用于提高基于特定结合反应的测试单元、尤其是免疫测试单元的动态测量范围的方法 |
US8372660B2 (en) | 2008-10-09 | 2013-02-12 | Actherm Inc | Quantitative analyzing method |
CN102187218B (zh) * | 2008-10-09 | 2014-02-12 | 红电医学科技股份有限公司 | 流体检测方法 |
CN102187218A (zh) * | 2008-10-09 | 2011-09-14 | 红电医学科技股份有限公司 | 流体检测方法 |
WO2010040250A1 (zh) * | 2008-10-09 | 2010-04-15 | 红电医学科技股份有限公司 | 流体检测方法 |
US8133718B2 (en) | 2008-10-17 | 2012-03-13 | Actherm Inc | Analytical strip and detecting method using the same |
US7964370B2 (en) | 2008-10-17 | 2011-06-21 | Actherm Inc | Analytical strip and detecting method using the same |
US8367015B2 (en) | 2009-03-23 | 2013-02-05 | Actherm Inc | Analytical strip and the manufacturing method thereof |
Also Published As
Publication number | Publication date |
---|---|
AU4222696A (en) | 1996-08-08 |
CA2166913A1 (en) | 1996-07-31 |
JPH08240591A (ja) | 1996-09-17 |
KR960029790A (ko) | 1996-08-17 |
EP0724157A2 (en) | 1996-07-31 |
ZA96357B (en) | 1996-08-01 |
US5569608A (en) | 1996-10-29 |
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