Phage display mediated immuno-PCR.
Phage display mediated immuno-PCR.
复制标题
噬菌体显示介导的免疫PCR。
DOI：
10.1093/nar/gkl260
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发表时间：
2006-05-08
影响因子：
14.9
通讯作者：
Zhang, Ji-Bin
中科院分区：
文献类型：
--
作者：
Guo, Yong-Chao;Zhou, Ya-Feng;Zhang, Xian-En;Zhang, Zhi-Ping;Qiao, Yan-Mei;Bi, Li-Jun;Wen, Ji-Kai;Liang, Mi-Fang;Zhang, Ji-Bin
免疫PCR（IPCR）因其超灵敏性，在免疫学研究和临床诊断中成为一种强有力的检测技术。在这里，我们介绍了一种新的策略，称为噬菌体展示介导的免疫PCR（PD-IPCR）。用重组噬菌体颗粒代替传统的IPCR中所需的单克隆抗体和DNA的化学键合，作为IPCR实验的现成试剂。表面展示的单链抗体和噬菌体DNA本身可分别直接作为检测抗体和PCR模板。本设计的目的是克服单链抗体检测灵敏度低的缺点，从而大大促进单链抗体在免疫分析中的真实的应用。以汉滩病毒核衣壳蛋白（NP）和朊病毒蛋白（PrP）为检测靶点，采用间接、夹心和实时荧光定量PCR方法进行了检测。与传统的酶联免疫吸附试验（ELISA）相比，检测灵敏度提高了1000- 10 000倍。该概念验证研究可作为一种新的模式，以开发一种易于操作，低成本和超灵敏的免疫测定方法，以广泛应用。
Immuno-PCR (IPCR) is a powerful detection technology in immunological study and clinical diagnosis due to its ultrasensitivity. Here we introduce a new strategy termed phage display mediated immuno-PCR (PD-IPCR). Instead of utilization of monoclonal antibody (mAb) and chemically bond DNA that required in the conventional IPCR, a recombinant phage particle is applied as a ready reagent for IPCR experiment. The surface displayed single chain variable fragment (scFv) and phage DNA themselves can directly serve as detection antibody and PCR template, respectively. The aim of the design is to overcome shortcoming of low detection sensitivity of scFv so as to largely facilitate the real application of scFv in immunoassay. The idea has been demonstrated by applying hantaan virus nucleocapsid protein (NP) and prion protein (PrP) as detection targets in three experimental protocols (indirect, sandwich and real-time PD-IPCR assays). The detection sensitivity was increased 1000- to 10 000-folds compared with conventional enzyme-linked immunosorbent assays (ELISAs). This proof-of-concept study may serve as a new model to develop an easy to operate, low cost and ultrasensitive immunoassay method for broad applications.
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影响因子：
14.9
作者：
HENDRICKSON, ER;TRUBY, TMH;EBERSOLE, RC
通讯作者：
EBERSOLE, RC
影响因子：
56.9
作者：
SANO, T;SMITH, CL;CANTOR, CR
通讯作者：
CANTOR, CR
影响因子：
7
作者：
Heid, CA;Stevens, J;Williams, PM
通讯作者：
Williams, PM
影响因子：
56.9
作者：
RUZICKA, V;MARZ, W;GROSS, W
通讯作者：
GROSS, W
DOI：
10.1016/s0006-291x(03)01364-0
发表时间：
2003-08-22
影响因子：
3.1
作者：
Adler, M;Wacker, R;Niemeyer, CM
通讯作者：
Niemeyer, CM