A reusable flow-through polymerase chain reaction instrument for the continuous monitoring of infectious biological agents
A reusable flow-through polymerase chain reaction instrument for the continuous monitoring of infectious biological agents
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DOI：
10.1021/ac034062u
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发表时间：
2003-07-15
影响因子：
7.4
通讯作者：
Marshall, GD
中科院分区：
文献类型：
--
作者：
Belgrader, P;Elkin, CJ;Marshall, GD
要持续监测环境中的传染病和相关生物战剂，就必须采用具有高度敏感性和特异性的切实可行、成本效益高的方法。在临床诊断中采用的一种相容方法是实时聚合酶链反应（PCR）分析。然而，由于商业PCR仪器中使用一次性消耗品，这种技术在环境监测中的实用性受到限制。这大大增加了机械复杂性，因为复杂的机器人机构必须补充一次性元件。另一种策略是开发一种自主监测系统，该系统由可重复使用的模块组成，这些模块易于与流通方案中的流体回路接口。降低的复杂性应增加可靠性，同时降低运营成本。在这份报告中，我们描述了一个可重复使用的，流通的PCR模块，功能作为一个组件，在这样的系统。该模块与炭疽杆菌基因组DNA进行了严格的评估，并证明了高的可重复性，灵敏度和效率，没有样品间携带的证据。
Continuous monitoring of the environment for infectious diseases and related biowarfare agents requires the implementation of practical cost-effective methodologies that are highly sensitive and specific. One compatible method employed in clinical diagnostics is real-time polymerase chain reaction (PCR) analysis. The utility of this technique for environmental monitoring is limited, however, by the utilization of single-use consumables in commercial PCR instruments. This greatly increases mechanical complexity, because sophisticated robotic mechanisms must replenish the disposable elements. An alternative strategy develops an autonomous monitoring system consisting of reusable modules that readily interface with fluidic circuitry in a flow-through scheme. The reduced complexity should increase reliability while decreasing operating costs. In this report, we describe a reusable, flow-through PCR module that functions as one component in such a system. This module was rigorously evaluated with Bacillus anthracis genomic DNA and demonstrated high repeatability, sensitivity, and efficiency, with no evidence of sample-to-sample carryover.