A semiautomated extraction protocol of HCV-RNA using Favorgen RNA extraction kit has been developed. The kit provided protocol was modified by replacing manual spin steps with vacuum filtration. The assay performance was evaluated by real-time qPCR based on Taqman technology. Assay linearity was confirmed with the serial dilutions of RTA (Turkey) containing 1 × (106, 105, 104, and 103) IU mL−1. Comparison of test results obtained by two extraction methods showed a good correlation (r = 0.95, n = 30) with detection limit of 102 IU mL−1. The semiautomated vacuum filtration based protocol demonstrated high throughput: 35 minutes for the extraction of a batch of 30 samples (150 µL each) with reduced labor, time, waste, and cost. Performance characteristics of semiautomated system make it suitable for use in diagnostic purpose and viral load determinations.
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机译：已经开发了使用Favorgen RNA提取试剂盒的HCV-RNA的半自动化提取方案。通过使用真空过滤代替手动旋转步骤，对试剂盒提供的方案进行了修改。通过基于Taqman技术的实时qPCR评估测定性能。用线性稀释的RTA（土耳其）证实了线性，该稀释液包含1×（10 6 ，10 5 ，10 4 和10 < sup> 3 ）IU mL -1 。两种提取方法获得的测试结果的比较显示出良好的相关性（r = 0.95，n = 30），检出限为10 2 IU mL -1 。基于半自动真空过滤的方案显示出高通量：提取一批批次的30个样品（每个150μL）需要35分钟，从而减少了劳动，时间，浪费和成本。半自动化系统的性能特征使其适合用于诊断目的和确定病毒载量。
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