Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay
Detection of human immunodeficiency virus type 1 by using the polymerase chain reaction and a time-resolved fluorescence-based hybridization assay
复制标题
使用聚合酶链式反应和基于时间分辨荧光的杂交测定法检测 1 型人类免疫缺陷病毒
DOI：
--
复制
发表时间：
1991
影响因子：
9.4
通讯作者：
M. Kwiatkowski
中科院分区：
文献类型：
--
作者：
P O Dahlen;A. Iitiä;G. Skagius;Å. Frostell;Michael Nunn;M. Kwiatkowski
聚合酶链式反应（PCR）在核酸诊断领域具有许多潜在的应用。特别是，它已成功应用于检测低拷贝数的病原体，例如 1 型人类免疫缺陷病毒。在这里，我们描述了一种基于时间分辨荧光的杂交测定，它与 PCR 相结合，为核酸检测提供了一种极其灵敏的方法。在此测定形式中，PCR 按标准程序运行，随后的杂交反应在溶液中使用两种寡核苷酸探针进行，一种是生物素化的，另一种是铕 (Eu3+) 标记的。然后将夹心杂交体收集到链霉亲和素包被的微量滴定孔上，并在时间分辨荧光计中测量结合的Eu3+。该测定快速、用户友好且定量，并且适合自动化。描述了该测定法在检测 1 型人类免疫缺陷病毒中的应用。
The polymerase chain reaction (PCR) has many potential applications in the field of nucleic acid diagnostics. In particular, it has been successfully applied to the detection of pathogens present in low copy numbers such as the human immunodeficiency virus type 1. Here we describe a time-resolved fluorescence-based hybridization assay which, combined with the PCR, offers an extremely sensitive method for the detection of nucleic acids. In this assay format, the PCR is run by standard procedures and the subsequent hybridization reaction is carried out in solution by using two oligonucleotide probes, one biotinylated and one labeled with europium (Eu3+). The sandwich hybrids are then collected onto a streptavidin-coated microtitration well, and the bound Eu3+ is measured in a time-resolved fluorometer. This assay is rapid, user friendly, and quantitative and lends itself to automation. The application of this assay to the detection of human immunodeficiency virus type 1 is described.
DOI：
10.1073/pnas.86.4.1173
发表时间：
1989-02-01
影响因子：
11.1
作者：
KWOH, DY;DAVIS, GR;GINGERAS, TR
通讯作者：
GINGERAS, TR
影响因子：
158.5
作者：
HO, DD;MOUDGIL, T;ALAM, M
通讯作者：
ALAM, M
影响因子：
158.5
作者：
EMBURY, SH;SCHARF, SJ;ERLICH, HA
通讯作者：
ERLICH, HA