Department of Virology, Federative Institute of Biology, CH Toulouse, France.
J Med Virol. 2012 Jun;84(6):906-11. doi: 10.1002/jmv.23281.
The aim of the study was to evaluate the MagNA Pure 96™ nucleic acid extraction system using clinical respiratory specimens for identifying viruses by qualitative real-time PCR assays. Three extraction methods were tested, that is, the MagNA Pure LC™, the COBAS Ampliprep™, and the MagNA Pure 96™ with 10-fold dilutions of an influenza A(H1N1)pdm09 sample. Two hundred thirty-nine respiratory specimens, 35 throat swabs, 164 nasopharyngeal specimens, and 40 broncho-alveolar fluids, were extracted with the MagNA Pure 96™ and the COBAS Ampliprep™ instruments. Forty COBAS Ampliprep™ positive samples were also tested. Real-time PCRs were used to identify influenza A and influenza A(H1N1)pdm09, rhinovirus, enterovirus, adenovirus, varicella zoster virus, cytomegalovirus, and herpes simplex virus. Similar results were obtained on RNA extracted from dilutions of influenza A(H1N1)pdm09 with the three systems: the MagNA Pure LC™, the COBAS Ampliprep™, and the MagNA Pure 96™. Data from clinical respiratory specimens extracted with the MagNA Pure 96™ and COBAS Ampliprep™ instruments were in 98.5% in agreement (P < 0.0001) for influenza A and influenza A(H1N1)pdm09. Data for rhinovirus were in 97.3% agreement (P < 0.0001) and in 96.8% agreement for enterovirus. They were in 100% agreement for adenovirus. Data for cytomegalovirus and HSV1-2 were in 95.2% agreement (P < 0.0001). The MagNA Pure 96™ instrument is easy-to-use, reliable, and has a high throughput for extracting total nucleic acid from respiratory specimens. These extracts are suitable for molecular diagnosis with any type of real-time PCR assay.
本研究旨在评估 MagNA Pure 96™ 核酸提取系统,使用临床呼吸道标本通过定性实时 PCR 检测来鉴定病毒。测试了三种提取方法,即 MagNA Pure LC™、COBAS Ampliprep™ 和 MagNA Pure 96™ 对流感 A(H1N1)pdm09 样本进行 10 倍稀释。用 MagNA Pure 96™ 和 COBAS Ampliprep™ 仪器提取了 239 份呼吸道标本、35 份咽喉拭子、164 份鼻咽标本和 40 份支气管肺泡液。还对 40 份 COBAS Ampliprep™ 阳性样本进行了检测。实时 PCR 用于鉴定流感 A 和流感 A(H1N1)pdm09、鼻病毒、肠道病毒、腺病毒、水痘带状疱疹病毒、巨细胞病毒和单纯疱疹病毒。从三种系统(MagNA Pure LC™、COBAS Ampliprep™ 和 MagNA Pure 96™)提取的流感 A(H1N1)pdm09 稀释液中获得了相似的 RNA 实时 PCR 结果:MagNA Pure LC™、COBAS Ampliprep™ 和 MagNA Pure 96™。用 MagNA Pure 96™ 和 COBAS Ampliprep™ 仪器提取的临床呼吸道标本的数据在流感 A 和流感 A(H1N1)pdm09 方面有 98.5%的一致性(P<0.0001)。鼻病毒数据的一致性为 97.3%(P<0.0001),肠道病毒数据的一致性为 96.8%。腺病毒数据的一致性为 100%。巨细胞病毒和 HSV1-2 的数据一致性为 95.2%(P<0.0001)。MagNA Pure 96™ 仪器易于使用、可靠,并且具有从呼吸道标本中提取总核酸的高通量。这些提取物适用于任何类型的实时 PCR 检测的分子诊断。