Petrik J, Pearson G J, Allain J P
Department of Haematology, University of Cambridge, UK.
J Virol Methods. 1997 Mar;64(2):147-59. doi: 10.1016/s0166-0934(96)02153-2.
An amplification procedure based on a semiautomated 60-sample RNA capture, including combined reverse transcription/polymerase chain reaction (RT-PCR) and nested PCR/Taqman amplicon detection. is described. It can be completed within a working day and is suitable for the development of a fully automated system. HCV RNA-specific capture is independent of the sequence variations as it targets the poly(U) tract commonly present at the 3'-end of the HCV genome (U-capture). The type specificity of the assay determined in a panel of 56 confirmed HCV antibody-positive samples (genotypes 1-6) was slightly better when compared to a commercial assay. The sensitivity evaluated on serial dilutions of representative samples was equal for genotypes 1, 2, 5, 6, or increased for genotypes 3 and 4 with the U-capture assay.
描述了一种基于半自动60样本RNA捕获的扩增程序，包括逆转录/聚合酶链反应（RT-PCR）和巢式PCR/Taqman扩增子检测相结合。该程序可在一个工作日内完成，适用于全自动系统的开发。丙型肝炎病毒（HCV）RNA特异性捕获不依赖于序列变异，因为它靶向HCV基因组3'端常见的聚（U）序列（U捕获）。在一组56份经确认的HCV抗体阳性样本（1-6型）中测定的该检测方法的类型特异性与商业检测方法相比略好。用U捕获检测法对代表性样本的系列稀释液进行评估时，1、2、5、6型的灵敏度相同，3型和4型的灵敏度有所提高。
