CN1321247A - 采用减少孔隙率以抑制迁移的测定 - Google Patents
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Abstract
一种测定分析物的装置,其中包括标记区,标记能在其中与分析物结合,标记区与捕获区相通,其中捕获区的孔隙尺寸使未与分析物结合的标记能迁移通过,而已与分析物结合的标记却不能迁移通过。在从标记区(孔隙尺寸大)向捕获区(孔隙尺寸小)迁移的过程中,未结合的标记能够进入和通过捕获区,而已结合的标记将在标记区和捕获区的连接处被捕获。该装置依赖于比分析物更小的标记,使游离的标记不受捕获区所阻止。该装置特别适合化验精子之类的分析物,精子比标记的抗体之类的标记大。
Description
技术领域
本发明涉及测量分析物的测定装置。特别是涉及采用机械方法,而不是采用常规的免疫捕获技术,在多孔材料中捕获分析物的装置。
技术背景
标准快速试验侧流装置的程式,在大约十年中仍没有改变。该装置一般包括硝化纤维素条。将试样施加到应用区,试样通过毛细管作用从应用区流过包含对所述对分析物特效的可见标记抗体的区域。游离的和已结合的标记继续向捕获区迁移,在其中对分析物专用的固定的抗体结合该分析物-标记复合物。游离标记(未结合的抗体)继续迁移,在捕获区留下分析物的独特信号。例如,在欧洲专利-A-0284232中公开了这些类型的侧流装置。叙述了包括在WO92/12428、欧洲专利-A-0613005、WO97/06439和美国专利-5,741,662中那些的许多种基本测定的方案。
然而,在所有情况下,都是由一种被固定的试剂作为捕获分析物-标记复合物的媒介,该试剂一般是对分析物特效的抗体。在许多方面这是不能令人满意的。
首先,制造质量控制是困难的。固相捕获膜一般是由硝化纤维素制造的,将抗体直接施加到膜上。然而,硝化纤维素制造是非均相的。因此,固相抗体的质量控制被限制于试验来自同一批非均相装置的统计试样,并假定整批都控制在规定的误差范围内。然而,众所周知,即使在一批膜中或在许多张膜中,膜的变化也是相当大的。
其次,它们的制造是比较麻烦的。将固定抗体施加到试条上需要一个与施加可移动标记抗体分开的单独步骤。捕获抗体可直接喷雾到硝化纤维素条上,但标记抗体则必须渗入随后采用搭接方法附着到硝化纤维素条上的材料中,以确保毛细流动。
第三,抗体是通过将溶液喷到膜上固定的。然而,有些抗体与膜结合得不牢固,有些仍然松弛地与固定的抗体相连。当溶剂前沿通过它时,这种半结合的或未结合的抗体可能变成能移动的抗体,造成在检测区标记的较少结合。如果该装置包括控制线,这将捕获本应在检测区被捕获的另一些标记。因此,依靠比较控制线和检测线之间颜色强度的化验如排卵预测试剂盒可能给出错误的结果。而且,通过喷雾施加不可避免地会引起扩散到膜中,导致检测信号更扩散及更不集中。
第四,该装置的灵敏度受其程式的限制。分析物和标记的抗体,随着它们通过膜迁移而发生反应,因此,调节流量能使标记的抗体在溶剂的前沿流动,使分析物-标记络合物的生成时间最长。然而,络合物在捕获抗体上通过的时间短,因此对试验设计及其操作特性施加了限制。反应时间短降低了灵敏度,也意味着需要高亲和力的捕获抗体。
最后,这些试验装置的储存期限往往受固定的捕获抗体在膜上随时间而破坏的限制。
本发明论述了现有技术装置的这些缺点,本发明并不采用固定的抗体去捕获分析物-标记复合物。
发明内容
本发明提供一种测定分析物的装置,它包括标记区,其中标记能与分析物结合,标记区与捕获区相通,其中捕获区孔隙的大小能使未与分析物结合的标记通过其迁移,而已与分析物结合的标记却不能通过其迁移。
在从标记区向捕获区迁移的过程中,未结合的标记能够进入并通过捕获区,因而,已结合的标记将在示踪区和捕获区的连接处被捕获,将捕获区入口处捕获的标记量与通过捕获区迁移的量进行比较,就能鉴定分析物的量-随着分析物浓度的增加,在标记区和捕获区连接处截留的标记量也增加。
显然,本发明依赖于比分析物更小的标记,使游离的标记不受捕获区的阻止。
该装置特别适合测定生物细胞之类的分析物,生物细胞比标记的抗体之类的标记大。优选的测定细胞是精子和细菌之类的微生物。
标记区是标记与分析物发生接触的区域。优选由纤维材料,例如高密度聚乙烯(HDPE)材料、粘结的聚酯纤维和玻璃纤维等制造的垫。与捕获区的孔隙尺寸相比,其孔隙尺寸应大到足以使分析物能比较自由地移动。
标记一般是能与有关分析物结合的抗体,并适合被标记。优选肉眼可见的标记,例如发荧光标记,或粒状的标记如胶体金(用肉眼看呈粉红色),或四溴荧光素之类的染色剂。应当理解,术语“抗体”可以包括多细胞系和单细胞系的抗体,以及抗体碎片(例如F(ab)2,Fc等),前提是保留必要的生物特性。
捕获区可由任何适宜的多孔材料制造,未结合的标记能通过这种多孔材料迁移,而已与分析物结合的标记却不能通过其迁移。这种要求反映在捕获区的孔隙大小。在一个实施方案中,捕获区是由名义孔隙尺寸约1-75μm,优选10-50μm,更优选20-35μm的HDPR制造的。在第二个实施方案中,捕获区是由具有名义孔隙尺寸约1-15μm,优选3-10μm,更优选5-8μm的硝化纤维素制造的。
在一些实施方案中,标记区和捕获区可以由单片多孔的材料制造,它包括孔隙尺寸减小的区域。例如,采用挤压或压实多孔材料的区域,可以将孔隙尺寸减小,以使由分析物结合的标记不能进入被压缩的区域,即制成捕获区。另外,将材料的孔隙堵塞一部分,也能达到同样的效果。
正如本领域人员所熟知的,多孔材料的名义孔隙尺寸可通过对坚固颗粒进行对比测试,即测定能通过该材料的球形颗粒的最大直径来确定。另外,材料的孔隙尺寸可通过测定其“泡点”来确定。泡点是强制空气通过(水)湿膜所需的压力,当根据颗粒滞留量测定时,泡点与孔隙的尺寸有关(虽然在极端压力和孔隙尺寸下,这种关系是较弱的)。泡点一般比颗粒滞留量容易测定,因此在鉴定孔隙尺寸时,优选测试泡点。
特别是,当采用本发明的装置检测和测量能移动的分析物时(例如能游动的精子或能游动的细菌),适宜的孔隙尺寸可通过日常试验根据经验来确定。
在优选的实施方案中,捕获区包括截留未与分析物结合的标记的区域(“标记控制”区)。该区一般包括固定在捕获区内的抗体,该抗体能与对对分析物特效的标记结合。通过捕获区的标记,并未在进入该区时被捕获,因此被截留在“标记控制”区内,并可在该区测定标记。例如,如果对分析物特效的标记是鼠的单细胞系抗体,那么捕获区则可包括包含固定的抗-鼠抗体的区域。因此,未结合的标记被截留在示踪区和捕获区的连接处,或在“标记控制”区。比较在这两处的示踪物量,就能鉴定分析物在原始试样中的量。
在另一种方案中,该装置可以利用在标记区内的二种分开的标记抗体,其中仅一种是对分析物特效的。该不能识别分析物的标记是“标记控制”区内的对抗体特效的标记。这种标记通过捕获区,被截留在“标记控制”区内,作为捕获区入口处对分析物的特效信号的比较标准。对分析物特效的标记不与“标记控制”区的抗体结合,并继续迁移。
标记区和捕获区的界面优选比捕获区的长度窄。在标记区和捕获区是由搭接的材料条制造时,它们之间较窄的界面可用无孔材料覆盖在大部分的搭接处上制成的。确保标记区和捕获区之间的界面较窄,能使分析物-标记复合物集中在标记区和捕获区的连接处,给出比较鲜明的信号。
该分析物优选为精子。标记优选能识别表面抗原,表面抗原存在于精子的主群上,而不是亚群上。因此,当可以采用任何表面抗原时,(例如,P34H(WO97/40836),SP-10(WO95/29188),也可参见欧洲专利-A-0387873),优选使用“通用抗原”如CD59。应当理解,在抗原对精子不特效的场合(即,例如CD59也存在在其它细胞类型上,),可能需要处理被分析的试样,从其中除去非精子细胞。阻止精子迁移的捕获区优选具有名义孔隙尺寸为5-8μm的硝化纤维素膜膜。可在分析之前处理精液试样以分离成能游动的和不能游动的细胞(例如,参见国际专利申请PCT/GB99/01929和PCT/GB99/02685)。可以使用本发明的装置,在比较分离后所得的结果可确定给定试样中能游动细胞和不能游动细胞的相对数目。本发明的装置可以包括在进入捕获区之前,分离能游动的精子和不能游动的精子的装置,以使化验之后出现三个信号—一个是标记接合不能游动的细胞的信号,一个是标记结合能游动的细胞的信号,一个是游离标记的信号。然而,在这种方法中,并不总是需要分离细胞,例如,在输精管结扎术的检验中,试验可以只指出能游动的或不能游动的精子的总数。通常分析包含精子的试样,而不是“纯”精液,而是被稀释的,可能是经过处理以除去了非精子细胞的试样。如果分析“纯”精液,一般需要使用对精子特效的标记,所以非精子细胞未被标记。
作为另一种方案,分析物可以是微生物体。微生物体可以是细菌,例如肠产毒性大肠杆菌(“ETEC”)[例如,见Levine(1987)传染病杂志(J.Infect.Dis)
155:377-289],对此可以采用任何适合标记的对ETEC特效的抗体作为标记,例如与金-配对的抗-CFA/I单细胞系。微生物体可以是酵母菌,例如念珠菌属。
在优选的实施方案中,为了促进毛细管的移动,可利用在标记区前和/或在捕获区后的毛细作用促进试样向捕获区迁移。
在本发明的一些实施方案中,可直接将试样施加到捕获区。在这个方案中,标记将从标记区通过捕获区迁移,在捕获区中遇到试样。标记将被已滞留在捕获区的分析物截留,未结合的标记物将继续迁移。
本发明的装置,可以简单便宜制成方便的试条或试片的形式。而且非常容易使用,例如可被家中用户使用。因此,本发明提供一种能在家中使用的测定装置,例如作为男性生育能力的基本鉴别装置。
附图简述
图1示出根据本发明的侧流动装置,图2示出采用本装置的剂量-响应曲线实验的结果。
实现本发明的方式实施例1-侧向流动检测的基本装置
图1所示的装置包括滤纸条(1)、包含金-标记的抗-精子标记抗体的第一种吸收剂垫(2)、接受含精子试样的第二种吸收剂垫(3)、硝化纤维素膜(4)和上部的毛细作用条(5)。在吸收剂垫(3)和硝化纤维素膜(4)之间,是乙酸酯薄条(6),该条除了在较窄的边界(7)处以外,能防止垫(3)与膜(4)之间的接触。膜(4)包括固定的抗体线(8),固定的抗体能与未结合的抗-精子标记抗体发生反应。
硝化纤维素膜(4)的尺寸为5mm×25mm,被固定在5mm×73mm的刚性塑料底片上。在膜(4)的一端,附着5mm×30mm的上部毛细作用条(5),使它们搭接5mm,在另一端固定尺寸为5mm×4mm的乙酸酯薄条(6),将尺寸为5mm×5mm的吸收剂垫(3),放在乙酸酯条(6)上,使其与硝化纤维素条(4)接触,其接触边界尺寸为5mm×1mm。将尺寸为5mm×5mm的吸收剂垫(2)附着,使其与垫(3)和条(6)邻接;将尺寸为20mm×5mm的滤纸(1)附着,使其在吸收剂垫(2)下面搭接2mm。
滤纸(1)是222号Aslstrom吸墨级滤纸。吸收剂垫(2)是HDPE复合材料,厚度为0.6mm,名义孔隙尺寸为99μm(英格兰,Sintair有限公司)。它用与40nm金颗粒(是用含5%茧蜜糖、0.1%Triton-X(Sigma)和1%BSA的纯化水稀释的)配对使用的单细胞系抗-CD59抗体(英国Bristol大学)溶液饱和,然后在真空下完全干燥。吸收剂垫(3)是由相同的HDPE材料制造的,但用1%的BAS和pH6.6的0.1%的Triton-X饱和,然后干燥。硝化纤维素膜(4)是Advanced MicroDevice的8μm硝化纤维素膜(CNPF-S1-L2-H50,批号为HF322228/731)。上部的毛细作用条(5)是由Whatman色谱纸制造的(分类号为3MM CHR,批号为3030640),在供料时未经处理。
控制抗体的细线(8)[Jackson’s AffiniPure山羊抗-鼠免疫球蛋白G(IgG)、可结晶的分段(FC)和专用分段(与人、牛和马的血清蛋白具有最小的交叉反应;编号115-005-071,批号36019),在2mM磷酸盐和0.017%BSA中稀释到0.02mg/ml]被固定在边界(7)和上部毛细作用条(5)之间的膜(4)上。
使用该装置时,将含精子的试样施加到吸收剂垫(3)上,并将滤纸(1)放在适宜的缓冲剂等中。缓冲剂通过滤纸(1)和吸收剂垫(2)迁移,使与金-配对的抗体与在垫(3)中的精子发生接触。当溶液通过垫(3)向边界(7)迁移时,抗体能与试样结合。被标记的精子不能通过硝化纤维素膜(4),这是因为其孔隙尺寸小,所以精子滞留在边界(7)附近。所以边界(7)起着“阻塞区”的作用,阻塞区捕获因为太大而不能通过膜(4)孔隙的物质。然而,未结合的与金-配对的抗体,继续通过膜迁移,直到其达到抗体线(8)为止,抗体线结合与金-配对的抗体。因此,在这一阶段,有二条可见的线—一条线是在阻塞区(7),那里通过与精子的结合,阻止了标记的迁移,另一条线是在线(8),那里通过与被固定的控制抗体的结合,阻止了迁移。实施例2-精子的定性试验
在试验中,试验二个试样-第一个试样是在HEPES缓冲剂中能游动的精子试样(是以间接方式从能生育的男人射精获得的)第二个试样就是HEPES缓冲剂。将每种试样的75μl分别放在二个装置的吸收剂垫(3)上,再将它们放在各自含有75μl HEPES的凹槽中。
在15分钟后,具有第一个试样的装置在膜(4)上出现二条清晰的红线,第一条在吸收剂垫(3)上,约1mm,第二条在线(8)上。然而,另一个装置只在线(8)上有一条红线。因此,该装置能用机械方法捕获“阻塞区”附近的精子。实施例3-精子的定量试验
在另一个试验中,通过间接方式从能生育的男人射精获得在HEPES缓冲剂中能游动的精子试样。采用计数室确定在HEPES中每毫升试样能游动的精子数,得到的结果是350万个。用HEPES稀释该试样,又得到每毫升分别具有200、100、50和25万个精子的四个试样。将这五个试样每个取75μl,分别放在五个装置的吸收剂垫(3)上,再将它们放在各自具有75μl HEPES的凹槽中。
在15分钟后,每个装置都在膜(4)上出现二条清晰的红线,第一条线在吸收剂垫(3)上,约1mm,第二条线在线(8)上。然而,正如在图2中清楚示出的,第一条线的强度,随着稀释作用使试样中的精子数降低而下降。对于对比试样看不到第一条线。因此该装置能够演示在阻塞区附近的计量响应捕获。另一些实施方案
应当理解,上面只通过实施例说明了本发明,还可以对本发明进行一些改进,这些改进也在本发明的范围和内容的范围内。
Claims (13)
1.一种测定分析物的装置,它包括标记区,标记在标记区内能与分析物结合,标记区与捕获区相通,其中捕获区的孔隙尺寸使未与分析物结合的标记能迁移通过,而与分析物结合的标记却不能迁移通过。
2.根据权利要求1的装置,其中的标记是能与相关的分析物结合的抗体。
3.根据权利要求2的装置,其中的标记是肉眼可见的。
4.根据权利要求3的装置,其中的标记是胶体金。
5.根据前述权利要求任一项的装置,其中的捕获区是硝化纤维素。
6.根据前述权利要求任一项的装置,其中的标记区和捕获区是由单片多孔材料制成的,这种多孔材料包含孔隙尺寸减小的区域。
7.根据前述权利要求任一项的装置,其中的捕获区包括截留未与分析物结合的标记的区域。
8.根据前述权利要求任一项的装置,其中标记区和捕获区的界面,与捕获区的长度相比是较窄的。
9.根据前述权利要求任一项的装置,其中的分析物是精子。
10.根据权利要求10的装置,其中的标记能识别CD59。
11.根据权利要求1-8任一项的装置,其中的分析物是微生物体。
12.根据前述权利要求任一项的装置,其中利用在标记区之前和/或在捕获区之后的毛细作用条促进试样向捕获区迁移。
13.根据前述权利要求任一项的装置,该装置是试条或试片的形式。
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101435823B (zh) * | 2007-11-12 | 2012-08-08 | 无锡中德伯尔生物技术有限公司 | 检测残留兽药的荧光微球免疫层析试纸条及其制备方法 |
CN103323590A (zh) * | 2013-06-08 | 2013-09-25 | 上海云泽生物科技有限公司 | 一种基于纤维膜捕集分离的定量检测装置及其检测方法 |
CN105004855A (zh) * | 2014-04-16 | 2015-10-28 | 万冰 | 一种侧向流动检测测试条 |
Families Citing this family (42)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7211255B2 (en) * | 1996-01-29 | 2007-05-01 | United States Of America The U.S. Environmental Protection Agency | Contraceptives based on SP22 and SP22 antibodies |
DE19927783A1 (de) * | 1999-06-18 | 2000-12-21 | Roche Diagnostics Gmbh | Element zur Bestimmung eines Analyts in einer Flüssigkeit, entsprechendes Bestimmungsverfahren unter Verwendung des Elementes sowie Kit zur Bestimmugn eines Analyts |
GB9925461D0 (en) * | 1999-10-27 | 1999-12-29 | Genosis Ltd | Assay device |
GB0008124D0 (en) * | 2000-04-03 | 2000-05-24 | Genosis Ltd | Capture assay |
US6837171B1 (en) | 2002-04-29 | 2005-01-04 | Palmer/Snyder Furniture Company | Lightweight table with unitized table top |
US8367013B2 (en) | 2001-12-24 | 2013-02-05 | Kimberly-Clark Worldwide, Inc. | Reading device, method, and system for conducting lateral flow assays |
US20030119203A1 (en) | 2001-12-24 | 2003-06-26 | Kimberly-Clark Worldwide, Inc. | Lateral flow assay devices and methods for conducting assays |
US7285424B2 (en) | 2002-08-27 | 2007-10-23 | Kimberly-Clark Worldwide, Inc. | Membrane-based assay devices |
EP1537416A1 (en) * | 2002-09-11 | 2005-06-08 | Lattec I/S | Device for analysing analyte compounds and use hereof |
US7781172B2 (en) | 2003-11-21 | 2010-08-24 | Kimberly-Clark Worldwide, Inc. | Method for extending the dynamic detection range of assay devices |
US7247500B2 (en) | 2002-12-19 | 2007-07-24 | Kimberly-Clark Worldwide, Inc. | Reduction of the hook effect in membrane-based assay devices |
WO2004081528A2 (en) * | 2003-03-10 | 2004-09-23 | Robinson Joseph R | Assay device and method |
US20040197819A1 (en) | 2003-04-03 | 2004-10-07 | Kimberly-Clark Worldwide, Inc. | Assay devices that utilize hollow particles |
US7851209B2 (en) | 2003-04-03 | 2010-12-14 | Kimberly-Clark Worldwide, Inc. | Reduction of the hook effect in assay devices |
US7393697B2 (en) | 2003-06-06 | 2008-07-01 | Advantage Diagnostics Corporation | Diagnostic test for analytes in a sample |
US7107936B2 (en) * | 2003-09-04 | 2006-09-19 | Mmi Genomics, Inc. | Device and method for animal tracking |
US7713748B2 (en) | 2003-11-21 | 2010-05-11 | Kimberly-Clark Worldwide, Inc. | Method of reducing the sensitivity of assay devices |
US7943395B2 (en) | 2003-11-21 | 2011-05-17 | Kimberly-Clark Worldwide, Inc. | Extension of the dynamic detection range of assay devices |
US20050112703A1 (en) | 2003-11-21 | 2005-05-26 | Kimberly-Clark Worldwide, Inc. | Membrane-based lateral flow assay devices that utilize phosphorescent detection |
US7943089B2 (en) | 2003-12-19 | 2011-05-17 | Kimberly-Clark Worldwide, Inc. | Laminated assay devices |
US20050221386A1 (en) * | 2004-02-17 | 2005-10-06 | Turner Nathan B | Chromatographic exclusion agglutination assay and methods of use thereof |
JP4616575B2 (ja) * | 2004-04-14 | 2011-01-19 | デンカ生研株式会社 | タンパク質の膜固定化方法 |
US20060019406A1 (en) * | 2004-07-23 | 2006-01-26 | Ning Wei | Lateral flow device for the detection of large pathogens |
US7442557B1 (en) * | 2004-10-22 | 2008-10-28 | Idexx Laboratories, Inc. | Bi-directional flow assay device with reagent bridge |
JP4547272B2 (ja) * | 2005-01-12 | 2010-09-22 | シスメックス株式会社 | イムノクロマトグラフィー用試験具 |
US7189522B2 (en) | 2005-03-11 | 2007-03-13 | Chembio Diagnostic Systems, Inc. | Dual path immunoassay device |
WO2006098804A2 (en) | 2005-03-11 | 2006-09-21 | Chembio Diagnostic Systems, Inc. | Dual path immunoassay device |
AU2006313611B2 (en) * | 2005-11-12 | 2013-01-31 | Platform Diagnostics Limited | Agglutination assay |
WO2007095530A2 (en) * | 2006-02-13 | 2007-08-23 | U.S. Environmental Protection Agency | Diagnostic kits to detect sp22 and sp22 antibodies |
US7704702B2 (en) * | 2006-08-10 | 2010-04-27 | Inverness Medical Switzerland Gmbh | Test strip for lateral flow assays |
GB2443694B (en) * | 2006-11-10 | 2011-09-14 | Platform Diagnostics Ltd | Analyte saturation assay, methods and kits and devices |
US8865454B2 (en) | 2007-03-22 | 2014-10-21 | Scandinavian Micro Biodevices Aps | Flow through system, flow through device and a method of performing a test |
US20090263905A1 (en) * | 2008-04-18 | 2009-10-22 | Kim Scheuringer | Detection test assembly for detecting the presence of a substance in a sample |
CN102084238B (zh) * | 2008-05-05 | 2016-06-01 | 洛斯阿拉莫斯国家安全有限责任公司 | 基于高度简化的侧向流动的核酸样品制备和被动流体流动控制 |
US20100022916A1 (en) | 2008-07-24 | 2010-01-28 | Javanbakhsh Esfandiari | Method and Apparatus for Collecting and Preparing Biological Samples for Testing |
US8828329B2 (en) | 2010-10-01 | 2014-09-09 | Church & Dwight, Co., Inc. | Electronic analyte assaying device |
US8603835B2 (en) | 2011-02-10 | 2013-12-10 | Chembio Diagnostic Systems, Inc. | Reduced step dual path immunoassay device and method |
SG11201608278WA (en) | 2014-04-02 | 2016-10-28 | Chembio Diagnostic Systems Inc | Immunoassay utilizing trapping conjugate |
US20160116466A1 (en) | 2014-10-27 | 2016-04-28 | Chembio Diagnostic Systems, Inc. | Rapid Screening Assay for Qualitative Detection of Multiple Febrile Illnesses |
US9981264B2 (en) | 2014-11-04 | 2018-05-29 | Grace Bio-Labs, Inc. | Nitrocellulose extrusion for porous film strips |
US20190145966A1 (en) * | 2016-04-12 | 2019-05-16 | Meje Ab | Membrane-based analytical device for bodily fluids |
JP6939106B2 (ja) * | 2017-06-09 | 2021-09-22 | 東洋紡株式会社 | イムノクロマト試験片およびキットおよび測定方法 |
Family Cites Families (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5763262A (en) * | 1986-09-18 | 1998-06-09 | Quidel Corporation | Immunodiagnostic device |
US5310650A (en) * | 1986-09-29 | 1994-05-10 | Abbott Laboratoires | Method and device for improved reaction kinetics in nucleic acid hybridizations |
US4857453A (en) * | 1987-04-07 | 1989-08-15 | Syntex (U.S.A.) Inc. | Immunoassay device |
DE291194T1 (de) * | 1987-04-27 | 1992-03-19 | Unilever N.V., Rotterdam | Immunoassays und vorrichtungen dafuer. |
JPS63305251A (ja) * | 1987-06-05 | 1988-12-13 | Dai Ichi Pure Chem Co Ltd | ラテツクス凝集反応を利用する免疫学的測定方法 |
US5244630A (en) * | 1988-04-22 | 1993-09-14 | Abbott Laboratories | Device for performing solid-phase diagnostic assay |
US5232834A (en) * | 1989-03-15 | 1993-08-03 | Fuso Pharmaceutical Industries, Ltd. | Anti-human sperm antibody, and its production and use |
AU636875B2 (en) * | 1989-03-17 | 1993-05-13 | Abbott Laboratories | Method and device for improved reaction kinetics in nucleic acid hybridizations |
US5087556A (en) * | 1989-05-17 | 1992-02-11 | Actimed Laboratories, Inc. | Method for quantitative analysis of body fluid constituents |
WO1994016329A1 (en) * | 1993-01-13 | 1994-07-21 | Abbott Laboratories | Methods for solid phase capture in immunoassays |
US5424193A (en) * | 1993-02-25 | 1995-06-13 | Quidel Corporation | Assays employing dyed microorganism labels |
ES2268706T3 (es) * | 1995-05-09 | 2007-03-16 | Beckman Coulter, Inc. | Dispositivos y procedimientos para separar componentes celulares de la sangre de la porcion liquida de la sangre. |
WO1996036878A1 (en) * | 1995-05-19 | 1996-11-21 | Universal Healthwatch, Inc. | Rapid self-contained assay format |
AU6720096A (en) * | 1995-08-09 | 1997-03-05 | Quidel Corporation | Test strip and method for one step lateral flow assay |
US5741662A (en) * | 1995-12-18 | 1998-04-21 | Quidel Corporation | Direct stain specific binding assays for microorganisms |
WO1997031259A1 (en) * | 1996-02-22 | 1997-08-28 | Dexall Biomedical Labs, Inc. | Non-captive substrate liquid phase immunoassay |
DE19629656A1 (de) * | 1996-07-23 | 1998-01-29 | Boehringer Mannheim Gmbh | Diagnostischer Testträger mit mehrschichtigem Testfeld und Verfahren zur Bestimmung von Analyt mit dessen Hilfe |
US6753189B1 (en) * | 1998-06-04 | 2004-06-22 | Mizuho Medy Co., Ltd. | Detection apparatus and method for the same |
-
1998
- 1998-10-02 GB GBGB9821526.2A patent/GB9821526D0/en not_active Ceased
-
1999
- 1999-10-01 JP JP2000574933A patent/JP4406510B2/ja not_active Expired - Fee Related
- 1999-10-01 CA CA002338796A patent/CA2338796A1/en not_active Abandoned
- 1999-10-01 GB GB0008683A patent/GB2345964B/en not_active Expired - Fee Related
- 1999-10-01 BR BR9914112-4A patent/BR9914112A/pt not_active Application Discontinuation
- 1999-10-01 AU AU61089/99A patent/AU759117B2/en not_active Ceased
- 1999-10-01 AT AT99947714T patent/ATE223054T1/de not_active IP Right Cessation
- 1999-10-01 CN CNB998116998A patent/CN1135391C/zh not_active Expired - Fee Related
- 1999-10-01 DE DE69902676T patent/DE69902676T2/de not_active Expired - Lifetime
- 1999-10-01 EP EP99947714A patent/EP1117997B1/en not_active Expired - Lifetime
- 1999-10-01 EP EP02078350A patent/EP1281968A3/en not_active Withdrawn
- 1999-10-01 ES ES99947714T patent/ES2182570T3/es not_active Expired - Lifetime
- 1999-10-01 WO PCT/GB1999/003249 patent/WO2000020866A1/en active IP Right Grant
-
2000
- 2000-09-08 US US09/658,148 patent/US6472226B1/en not_active Expired - Lifetime
-
2002
- 2002-05-13 US US10/145,170 patent/US7211403B2/en not_active Expired - Fee Related
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101435823B (zh) * | 2007-11-12 | 2012-08-08 | 无锡中德伯尔生物技术有限公司 | 检测残留兽药的荧光微球免疫层析试纸条及其制备方法 |
CN103323590A (zh) * | 2013-06-08 | 2013-09-25 | 上海云泽生物科技有限公司 | 一种基于纤维膜捕集分离的定量检测装置及其检测方法 |
CN105004855A (zh) * | 2014-04-16 | 2015-10-28 | 万冰 | 一种侧向流动检测测试条 |
CN105004855B (zh) * | 2014-04-16 | 2017-05-31 | 万冰 | 一种侧向流动检测测试条 |
Also Published As
Publication number | Publication date |
---|---|
CA2338796A1 (en) | 2000-04-13 |
GB9821526D0 (en) | 1998-11-25 |
DE69902676D1 (de) | 2002-10-02 |
WO2000020866A1 (en) | 2000-04-13 |
ES2182570T3 (es) | 2003-03-01 |
EP1117997B1 (en) | 2002-08-28 |
EP1281968A2 (en) | 2003-02-05 |
DE69902676T2 (de) | 2003-04-24 |
ATE223054T1 (de) | 2002-09-15 |
GB0008683D0 (en) | 2000-05-31 |
BR9914112A (pt) | 2001-06-12 |
AU759117B2 (en) | 2003-04-03 |
US6472226B1 (en) | 2002-10-29 |
GB2345964A (en) | 2000-07-26 |
US7211403B2 (en) | 2007-05-01 |
JP4406510B2 (ja) | 2010-01-27 |
CN1135391C (zh) | 2004-01-21 |
EP1281968A3 (en) | 2003-05-14 |
EP1117997A1 (en) | 2001-07-25 |
US20020127614A1 (en) | 2002-09-12 |
GB2345964B (en) | 2000-12-06 |
JP2002526774A (ja) | 2002-08-20 |
AU6108999A (en) | 2000-04-26 |
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