Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network.
Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network.
复制标题
DOI：
10.1021/ac500872j
复制
发表时间：
2014-07-01
影响因子：
7.4
通讯作者：
Yager, Paul
中科院分区：
文献类型：
--
作者：
Fridley, Gina E.;Le, Huy;Yager, Paul
我们已经展示了基于图案化的干燥试剂的受控再水化的多步骤二维纸网络免疫测定。先前的工作已经表明，信号增强提高了2维纸网络测定中的检测极限，但是直到现在，试剂仅以湿的或干的形式包括在放置在测定装置上游端的单独的缀合物垫中。湿试剂对于护理点来说并不理想，因为它们必须冷藏，并且通常限制自动化并且需要更多的用户步骤。结合物垫允许干燥，但不能在再水化时提供对试剂分布的任何控制，并且当垫不均匀地接触测定膜时，结合物垫可能是错误的来源。此外，每种试剂在单独的垫上干燥，当实施需要几种不同试剂的多步测定时增加了制造复杂性。相反，我们的新方法允许直接在纸膜内的图案化试剂储存库进行一致的、受控的再水化。在该试验演示中，在试验装置的不同区域中对四种单独的试剂进行了图案化：用于抗原检测的金抗体结合物和三种不同的信号增强组分，这些组分必须在临用前混合。为了显示图案化和干燥试剂直接到纸装置上用于干燥试剂储存和随后的受控释放的可行性，我们用疟疾抗原恶性疟原虫富组氨酸蛋白2（PfHRP 2）作为靶分析物的实例测试了该装置。在该演示中，信号增强步骤使可见信号增加约3倍，并使分析检测限降低2.75倍。
We have demonstrated a multistep 2-dimensional paper network immunoassay based on controlled rehydration of patterned, dried reagents. Previous work has shown that signal enhancement improves the limit of detection in 2-dimensional paper network assays, but until now, reagents have only been included as wet or dried in separate conjugate pads placed at the upstream end of the assay device. Wet reagents are not ideal for point-of-care because they must be refrigerated and typically limit automation and require more user steps. Conjugate pads allow drying but do not offer any control of the reagent distribution upon rehydration and can be a source of error when pads do not contact the assay membrane uniformly. Furthermore, each reagent is dried on a separate pad, increasing the fabrication complexity when implementing multistep assays that require several different reagents. Conversely, our novel method allows for consistent, controlled rehydration from patterned reagent storage depots directly within the paper membrane. In this assay demonstration, four separate reagents were patterned in different regions of the assay device: a gold-antibody conjugate used for antigen detection and three different signal enhancement components that must not be mixed until immediately before use. To show the viability of patterning and drying reagents directly onto a paper device for dry reagent storage and subsequent controlled release, we tested this device with the malaria antigen Plasmodium falciparum histidine-rich protein 2 (PfHRP2) as an example of target analyte. In this demonstration, the signal enhancement step increases the visible signal by roughly 3-fold and decreases the analytical limit of detection by 2.75-fold.
登录
查看更多内容
评估PFHRP2和基于PLDH的快速诊断测试，以诊断2名非洲儿童种群的严重疟疾诊断。
DOI：
10.1093/cid/cir143
发表时间：
2011-05
期刊：
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America
影响因子：
--
作者：
Hendriksen IC;Mtove G;Pedro AJ;Gomes E;Silamut K;Lee SJ;Mwambuli A;Gesase S;Reyburn H;Day NP;White NJ;von Seidlein L;Dondorp AM
通讯作者：
Dondorp AM
影响因子：
9.4
作者：
Cazacu, AC;Greer, J;Demmler, GJ
通讯作者：
Demmler, GJ
影响因子：
7.4
作者：
Hossain, S. M. Zakir;Luckham, Roger E.;Brennan, John D.
通讯作者：
Brennan, John D.
影响因子：
3
作者：
Butterworth AS;Robertson AJ;Ho MF;Gatton ML;McCarthy JS;Trenholme KR
通讯作者：
Trenholme KR
影响因子：
6.1
作者：
Govindarajan, A. V.;Ramachandran, S.;Boehringer, K. F.
通讯作者：
Boehringer, K. F.