Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02114, USA.
Nat Commun. 2013;4:1752. doi: 10.1038/ncomms2745.
The task of rapidly identifying patients infected with Mycobacterium tuberculosis in resource-constrained environments remains a challenge. A sensitive and robust platform that does not require bacterial isolation or culture is critical in making informed diagnostic and therapeutic decisions. Here we introduce a platform for the detection of nucleic acids based on a magnetic barcoding strategy. PCR-amplified mycobacterial genes are sequence-specifically captured on microspheres, labelled by magnetic nanoprobes and detected by nuclear magnetic resonance. All components are integrated into a single, small fluidic cartridge for streamlined on-chip operation. We use this platform to detect M. tuberculosis and identify drug-resistance strains from mechanically processed sputum samples within 2.5 h. The specificity of the assay is confirmed by detecting a panel of clinically relevant non-M. tuberculosis bacteria, and the clinical utility is demonstrated by the measurements in M. tuberculosis-positive patient specimens. Combined with portable systems, the magnetic barcode assay holds promise to become a sensitive, high-throughput and low-cost platform for point-of-care diagnostics.
在资源有限的环境中快速识别感染结核分枝杆菌的患者仍然是一项挑战。一个不依赖于细菌分离或培养的灵敏且稳健的平台，对于做出明智的诊断和治疗决策至关重要。在这里，我们介绍了一种基于磁条码策略的核酸检测平台。通过聚合酶链反应（PCR）扩增的分枝杆菌基因在微球上被序列特异性捕获，用磁性纳米探针标记，并通过核磁共振检测。所有组件都集成到一个单一的、小型的流体盒中，以实现芯片上的流畅操作。我们使用该平台从机械处理的痰样本中在 2.5 小时内检测结核分枝杆菌并鉴定耐药菌株。该检测方法的特异性通过检测一组临床相关的非结核分枝杆菌细菌得到证实，其临床实用性通过对结核分枝杆菌阳性患者样本的测量得到证实。与便携式系统相结合，磁条码检测法有望成为一种灵敏、高通量、低成本的即时诊断平台。
