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磁珠法核酸纯化试剂盒:粪便标本中的临床应用及性能评估

Magnetic bead-based nucleic acid purification kit: Clinical application and performance evaluation in stool specimens.

作者信息

Yoon Jihoon G, Kang Jin Seok, Hwang Seung Yong, Song Jaewoo, Jeong Seok Hoon

机构信息

Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, Republic of Korea.

Molecular Diagnostics Team, SK Telecom, Sungnam-si, Gyeonggi-do, Republic of Korea; Department of Molecular and Life Science, Hanyang University, Ansan, Gyeonggi-do, Republic of Korea.

出版信息

J Microbiol Methods. 2016 May;124:62-8. doi: 10.1016/j.mimet.2016.03.011. Epub 2016 Mar 24.

Abstract

Two different methods - the semi-automated magnetic bead-based kit (SK, Stool DNA/RNA Purification kit®) and the manual membrane column-based kit (QS, QIAamp® DNA Stool Mini kit) - for purifying nucleic acids from clinical stool samples were compared and evaluated. The SK kit was more user-friendly than QS due to the reduced manual processing, partial automation, and short turnaround time with half cost. Furthermore, SK produced high yields in both DNA and RNA extractions but poor purity in RNA extraction. In the assessment of rotavirus and Clostridium difficile infection, both kits had equivalent or more sensitive performance compared with the standard method. Although SK showed some interference and inhibition in nucleic acid extraction, the performance, including the repeatability, linearity, analytical sensitivity, and matrix effect, was sufficient for routine clinical use.

摘要

比较和评估了两种从临床粪便样本中纯化核酸的不同方法——基于半自动磁珠的试剂盒(SK,粪便DNA/RNA纯化试剂盒®)和基于手动膜柱的试剂盒(QS,QIAamp® DNA粪便微量试剂盒)。由于手动操作减少、部分自动化以及周转时间短且成本减半,SK试剂盒比QS试剂盒更便于用户使用。此外,SK在DNA和RNA提取中均产生高产率,但RNA提取的纯度较差。在轮状病毒和艰难梭菌感染的评估中,与标准方法相比,两种试剂盒具有相当或更敏感的性能。尽管SK在核酸提取中表现出一些干扰和抑制作用,但其性能,包括重复性、线性、分析灵敏度和基质效应,足以满足常规临床使用。

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