O'Meara D, Yun Z, Sönnerborg A, Lundeberg J
Department of Biochemistry and Biotechnology, Royal Institute of Technology (KTH), Stockholm, Sweden.
J Clin Microbiol. 1998 Sep;36(9):2454-9. doi: 10.1128/JCM.36.9.2454-2459.1998.
A novel method for direct capture of hepatitis C virus (HCV) RNA from clinical samples has been developed. This approach takes advantage of the cooperative interactions between adjacently hybridized oligonucleotides. Here, this cooperative effect was combined with solid-phase technology, whereby a capture probe was covalently coupled to magnetic beads and a second probe, which anneals adjacent to the capture probe site, was prehybridized in solution to the target. When these contiguously hybridized probes were used for the extraction of HCV RNA from clinical samples, the capture efficiency was increased up to 25-fold in comparison to capture with a single probe. The applicability of this sample preparation assay was further investigated by performing a comparative study with both a conventional guanidinium extraction method and a commercial quantitative assay.
一种从临床样本中直接捕获丙型肝炎病毒(HCV)RNA的新方法已经开发出来。这种方法利用了相邻杂交寡核苷酸之间的协同相互作用。在此,这种协同效应与固相技术相结合,即捕获探针共价偶联到磁珠上,另一个与捕获探针位点相邻退火的探针在溶液中与靶标预杂交。当这些连续杂交的探针用于从临床样本中提取HCV RNA时,与使用单一探针捕获相比,捕获效率提高了25倍。通过与传统的胍提取方法和商业定量测定法进行比较研究,进一步研究了这种样本制备测定法的适用性。