Sefers Susan E, Rickmyre Jamie, Blackman Amondrea, Li Haijing, Edwards Kathryn, Tang Yi-Wei
Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
J Clin Virol. 2006 Feb;35(2):141-6. doi: 10.1016/j.jcv.2005.05.011. Epub 2005 Jul 21.
Nucleic acid preparation from a variety of clinical specimens requires efficient target recovery and amplification inhibitor removal and is critical for successful molecular diagnosis. The QIAamp MinElute Virus kit (Qiagen Inc., Valencia, CA) was compared to the two existing methods currently used in our laboratory, IsoQuick (Orca Research Inc., Bothell, WA) for DNA extraction and RNAzol B (Leedo Laboratories Inc., Houston, TX) for RNA extraction, of viral nucleic acids.
A total of 150 clinical specimens, including cerebrospinal fluid (CSF) and nasopharyngeal swabs (NPS), were used to determine the extraction efficiency of the MinElute compared to the other two methods. Nucleic acid recovery, hands-on time, turn-around-time and cost were compared across all kits.
There was complete concordance between the MinElute and IsoQuick/RNAzol kits when herpes simplex virus (HSV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), influenza A virus or enteroviruses were detected using a colorimetric microtiter plate PCR system. The kits were equivalent in their ability to detect either DNA or RNA with superior ability to recover a high quality and quantity of RNA. With the potential to process larger specimen volumes, the MinElute kit can significantly shorten processing time from 2h to 50-55min.
Although relatively high test kit costs were noted, the MinElute kit provides another rapid and user-friendly specimen processing tool in the diagnostic molecular microbiology laboratory.
从各种临床标本中制备核酸需要高效回收靶标并去除扩增抑制剂，这对于成功进行分子诊断至关重要。将QIAamp MinElute病毒试剂盒（Qiagen公司，加利福尼亚州瓦伦西亚）与我们实验室目前使用的两种现有方法进行了比较，即用于DNA提取的IsoQuick（Orca Research公司，华盛顿州博塞尔）和用于RNA提取的RNAzol B（Leedo Laboratories公司，得克萨斯州休斯顿），以提取病毒核酸。
总共150份临床标本，包括脑脊液（CSF）和鼻咽拭子（NPS），用于确定MinElute与其他两种方法相比的提取效率。对所有试剂盒的核酸回收率、实际操作时间、周转时间和成本进行了比较。
当使用比色微量滴定板PCR系统检测单纯疱疹病毒（HSV）、爱泼斯坦-巴尔病毒（EBV）、水痘-带状疱疹病毒（VZV）、甲型流感病毒或肠道病毒时，MinElute与IsoQuick/RNAzol试剂盒之间的检测结果完全一致。这些试剂盒在检测DNA或RNA方面能力相当，在回收高质量和高数量RNA方面能力更强。由于有可能处理更大体积样本，MinElute试剂盒可将处理时间从2小时显著缩短至50 - 55分钟。
尽管注意到试剂盒成本相对较高，但MinElute试剂盒为诊断分子微生物学实验室提供了另一种快速且用户友好的标本处理工具。
