The quality of extracted DNA is crucial for obtaining accurate results, and automated extraction methods are faster and more effective than manual extraction. The novel Real-Prep automated extraction system has not yet been verified by direct comparisons to existing methods. In this study, we compared it to manual extraction and the most common automated extraction method used in Korea, the Nextractor system. From August to December 2018, 238 specimens, including sputum, bronchial washing, pericardial fluid, bronchial aspiration, pleural fluid, and closed pus samples, were collected and examined at Yeungnam University Hospital. After decontamination, smear microscopy, and culturing, DNA was extracted using the three methods. The DNA extraction efficiency (total amount of DNA [ng]/input specimen volume [μL]) and purity (A260/280 ratio), which indicates the presence of contaminants, were compared. Real-time PCR tests were conducted using the DNA extracted by each method. The cycle threshold, which is inversely related to the initial amount of mycobacterial DNA, and the percentage agreement between the PCR results of the three methods were evaluated. The PCR results were compared for sensitivity and specificity. Our study revealed that the DNA extraction efficiency of the Real-Prep system was higher than that of manual extraction. There was no significant difference in DNA purity between the methods, and the percentage agreement for Mycobacterium tuberculosis and non-tuberculous mycobacteria among all three methods was nearly 100%. Sensitivity for detecting non-tuberculous mycobacteria was higher for the Real-Prep system (71%, 34/48) than for the manual method (56%, 25/48). The performance of the Real-Prep system was similar to that of the Nextractor system and superior to that of manual extraction. The Real-Prep system, a new automated nucleic acid extraction device, has a clear benefit because of its relative speed and low hands-on time. Therefore, the Real-Prep system is a useful substitute for manual DNA extraction, which has the potential to reduce workloads in laboratories and as a sensitive non-tuberculous mycobacteria detection method throughout the world.

中文翻译：

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用于提取核酸以检测分枝杆菌感染的自动Real-Prep系统的临床评估。

提取的DNA的质量对于获得准确的结果至关重要，并且自动提取方法比手动提取更快，更有效。新型Real-Prep自动提取系统尚未通过与现有方法的直接比较得到验证。在这项研究中，我们将其与手动提取以及韩国使用的最常见的自动提取方法Nextractor系统进行了比较。从2018年8月至2018年12月，在Yeungnam University Hospital收集并检查了238个标本，包括痰液，支气管冲洗液，心包积液，支气管抽吸液，胸膜积液和闭合性脓液样本。净化，涂片显微镜检查和培养后，使用三种方法提取DNA。DNA提取效率（DNA总量[ng] /输入样品体积[μL]）和纯度（A260 / 280比），比较表明存在污染物。使用每种方法提取的DNA进行实时PCR测试。循环阈值与分枝杆菌DNA的初始量成反比，并且评估了三种方法的PCR结果之间的百分比一致性。比较PCR结果的敏感性和特异性。我们的研究表明，Real-Prep系统的DNA提取效率要高于手工提取。两种方法之间的DNA纯度没有显着差异，三种方法中结核分枝杆菌和非结核分枝杆菌的百分比一致性接近100％。Real-Prep系统检测非结核分枝杆菌的灵敏度（71％，34/48）高于手动方法（56％，25/48）。Real-Prep系统的性能类似于Nextractor系统，并且优于手动提取。Real-Prep系统是一种新型的自动核酸提取设备，其相对速度快且动手时间短，因此具有明显的优势。因此，Real-Prep系统是替代人工DNA提取的有用替代方法，具有减少实验室工作量的潜力，并且是全世界范围内灵敏的非结核分枝杆菌检测方法。