Multiplex detection of bacteria on an integrated centrifugal disk using bead-beating lysis and loop-mediated amplification.
Multiplex detection of bacteria on an integrated centrifugal disk using bead-beating lysis and loop-mediated amplification.
复制标题
使用珠打裂解和环介导扩增对集成离心盘上的细菌进行多重检测
DOI：
10.1038/s41598-017-01415-x
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发表时间：
2017-05-03
影响因子：
4.6
通讯作者：
Xing W
中科院分区：
文献类型：
--
作者：
Yan H;Zhu Y;Zhang Y;Wang L;Chen J;Lu Y;Xu Y;Xing W
虽然基于培养的细菌鉴定是诊断传染病的金标准，但它很耗时。分子诊断和微流控技术的最新进展为快速检测细菌开辟了新的途径。在这里，我们描述了一个离心微流控芯片检测细菌通过集成细胞裂解，澄清，环介导扩增（LAMP）。该芯片的主要优点如下。首先，细菌裂解创新地通过旋转一对磁体产生珠撞击而实现，同时在裂解过程中保持芯片静止，这简化了芯片设计，因为不需要额外的阀。其次，芯片上测定时间短（70分钟内），这在紧急情况下具有竞争力。第三，分析结果可以通过使用荧光检测器或肉眼进行解释，使其在许多领域，特别是资源有限的地区具有通用性。采用凝胶电泳法对6种细菌的芯片检测限进行了评价，结果与实验室LAMP方法相似。该芯片可用于快速、灵敏、准确和自动化的细菌检测，为简化传染病的分子诊断提供了一种有前途的替代方案。
Although culture-based identification of bacteria is the gold-standard for the diagnosis of infectious diseases, it is time consuming. Recent advances in molecular diagnostics and microfluidic technologies have opened up new avenues for rapid detection of bacteria. Here, we describe a centrifugal-microfluidic chip for the detection of bacteria by integrating the cell lysis, clarification, and loop-mediated amplification (LAMP). The major advantages of this chip are as follows. Firstly, bacteria lysis was innovatively achieved by rotating a pair of magnets to generate bead-beating while the chip was kept stationary during lysis, which simplified the chip design because no additional valve was needed. Secondly, the on-chip assay time was short (within 70 min), which was competitive in emergency situations. Thirdly, results of the analysis can be interpreted by using a fluorescence detector or by the naked-eye, making it versatile in many areas, especially the resource-limited areas. The on-chip limits of detection of six types of bacteria were valued by gel electrophoresis, showing the similar results compared to the bench-top LAMP protocol. This chip can be used for rapid, sensitive, accurate and automated detection of bacteria, offering a promising alternative for simplifying the molecular diagnostics of infectious diseases.