Centre for Drugs and Diagnostics, Liverpool School of Tropical Medicine, Liverpool, UK.
Malawi Liverpool Wellcome Trust, Blantyre, Malawi.
J Appl Microbiol. 2022 Nov;133(5):3191-3200. doi: 10.1111/jam.15755. Epub 2022 Aug 23.
The environment is increasingly recognized as an important reservoir of antimicrobial resistance genes (ARGs), which can be identified using molecular platforms. Yet, environmental surveillance remains an underutilised tool as there is no agreement on the best strategy for sample processing. We aim to develop a low-cost extraction method independent to commercial kits or reagents.
We present a novel, magnetic bead-based method for the isolation of ARGs from river water named MagnaExtract. We present this with analytic limit of detection as well as a case study in Southern Malawi. Here we compare the DNA yield from MagnaExtract with commercially available QIAGEN kits and the crude boil and spin method, using a high-resolution melt analysis PCR panel designed for the detection of third-generation cephalosporin and carbapenem-resistant genes from 98 water samples.
The MagnaExtract method is comparable, and in some instance's superior to commercially available kits for the isolation of ARGs from river water samples.
The MagnaExtract approach offers a simple, affordable, high yielding extraction method that could be used for the detection of ARGs from river water samples in surveillance campaigns in East Africa.
环境越来越被认为是抗菌药物耐药基因(ARGs)的重要储存库,可以使用分子平台来识别。然而,环境监测仍然是一个未被充分利用的工具,因为对于最佳的样本处理策略尚未达成共识。我们旨在开发一种独立于商用试剂盒或试剂的低成本提取方法。
我们提出了一种从河水中分离 ARGs 的新型磁性珠基方法,称为 MagnaExtract。我们提供了该方法的分析检测限,以及在马拉维南部的案例研究。在这里,我们使用针对第三代头孢菌素和碳青霉烯类耐药基因设计的高分辨率熔解分析 PCR 面板,比较了 MagnaExtract 与市售的 QIAGEN 试剂盒以及粗煮沸和旋转法从 98 个水样中提取的 DNA 产量。
MagnaExtract 方法可与市售试剂盒相媲美,在某些情况下,从河水样本中分离 ARGs 的效果优于市售试剂盒。
MagnaExtract 方法提供了一种简单、经济、高产量的提取方法,可用于东非的河流水样中 ARGs 的检测。