Yagmur Gulhan, Altun Hatice Uludag, Gökahmetoglu Selma, Basok Ela
Council of Forensic Medicine, Postmortem Microbiology, Istanbul, Turkey.
Turgut Ozal University Faculty of Medicine, Department of Medical Microbiology, Ankara, Turkey.
Infez Med. 2015 Sep;23(3):247-52.
In the diagnosis and monitoring of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, it is important to use methods that can provide rapid and reliable results. The present study aimed to compare the automated and manual extraction methods during the nucleic acid isolation phase for HBV-DNA and HCV-RNA assays. The study included 93 serum samples, 49 of which were for the HBV-DNA assay and 44 for the HCV-RNA assay. DNA and RNA isolation from the samples was performed manually with a "QIAmpMin Elute Kit" (Qiagen, Germany) and the automated isolation system, NucliSens easyMAG (BioMérieux, France). All the extraction products were amplified using the iCycler device (Bio-Rad, USA). With both methods, compliance was found in 21 (42.8%) samples in the HBV-DNA assay; nine (18.3%) samples had a higher amount of viral nucleic acid with the manual method, whereas 19 samples (38.7%) were found to have a higher amount of nucleic acid with the automated system. For the HCV-RNA assay, total compliance was found in 31 (70.4%) samples; 12 (27.2%) samples had a higher amount of viral nucleic acid with the manual method whereas one sample (2.2%) was found to have a higher amount of nucleic acid with the automated system. It was concluded that the NucliSens easyMAG automated isolation system can be used with confidence for nucleic acid extraction due to its higher sensitivity, providing results in a shorter time, and assured standardization.
在乙型肝炎病毒（HBV）和丙型肝炎病毒（HCV）感染的诊断和监测中，使用能够提供快速可靠结果的方法非常重要。本研究旨在比较HBV-DNA和HCV-RNA检测中核酸提取阶段的自动化和手动提取方法。该研究纳入了93份血清样本，其中49份用于HBV-DNA检测，44份用于HCV-RNA检测。使用“QIAmpMin Elute试剂盒”（德国Qiagen公司）和自动化提取系统NucliSens easyMAG（法国生物梅里埃公司）对样本进行DNA和RNA的手动提取。所有提取产物均使用iCycler仪器（美国伯乐公司）进行扩增。在HBV-DNA检测中，两种方法均在21份（42.8%）样本中获得符合结果；9份（18.3%）样本手动提取方法获得的病毒核酸量更高，而19份样本（38.7%）自动化系统提取的核酸量更高。对于HCV-RNA检测，31份（70.4%）样本获得完全符合结果；12份（27.2%）样本手动提取方法获得的病毒核酸量更高，而1份样本（2.2%）自动化系统提取的核酸量更高。研究得出结论，NucliSens easyMAG自动化提取系统因其更高的灵敏度、更短的出结果时间和可靠的标准化，可放心用于核酸提取。
