Detection of human immunodeficiency virus type 1 DNA by polymerase chain reaction amplification and capture hybridization in microtiter wells.
Detection of human immunodeficiency virus type 1 DNA by polymerase chain reaction amplification and capture hybridization in microtiter wells.
复制标题
通过聚合酶链式反应扩增和微量滴定孔中的捕获杂交检测人类免疫缺陷病毒 1 型 DNA。
DOI：
10.1128/jcm.29.3.638-641.1991
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发表时间：
1991
影响因子：
9.4
通讯作者：
Manak,MM
中科院分区：
文献类型：
--
作者：
Keller,GH;Huang,DP;Manak,MM
我们开发了一种改进的基于微滴度的检测聚合酶链反应（PCR）扩增DNA序列的方法。用于引物PCR的合成DNA序列在其5'端标记生物素，以便特定的PCR产物标记生物素。扩增后，将PCR产物的等分物变性并与固定在微滴孔中的捕获DNA序列杂交。捕获序列与引物之间的部分序列互补，因此只捕获延伸引物。用链亲和素过氧化物酶偶联物和四甲基联苯胺底物比色法检测捕获的PCR产物。
We developed an improved microtiter-based assay for the detection of polymerase chain reaction (PCR)-amplified DNA sequences. The synthetic DNA sequences used to prime the PCR were labeled with biotin at their 5' ends so that the specific PCR product was labeled with biotin. Following amplification, an aliquot of the PCR product was denatured and hybridized to a capture DNA sequence immobilized in a microtiter well. The capture sequence was complementary to a portion of the sequence between the primers, so that only extended primers were captured. The captured PCR product was detected colorimetrically by using a streptavidin-peroxidase conjugate and tetramethylbenzidine substrate.