微流控振荡流PCR快速检测单增李斯特氏菌

摘要

一种高通量振荡流PCR微流控技术已被成功开发,借此来快速检测食品致病菌-单增李斯特氏菌(L.monocytogenes).该PCR微流控装置主要由基于LabView的温度控制与采集系统、三个铜加热块以及聚四氟乙烯(PTFE)毛细管等构成.在该微流控装置上,三个铜块维持PCR反应所需要的三个温度,而包埋在铜块沟槽中的PTFE毛细管作为PCR的反应通道.毛细管通道中的PCR溶液通过精密注射泵驱动实现往复振荡,从而实现基于振荡流PCR的DNA快速扩增.当流速为300 μL/min时,135 bp DNA基因片段能在14 min内成功扩增,L.monocytogenes检测极限为10 cfu/mL.目前开发的高通量振荡流PCR系统能同时检测水、牛奶、香焦以及香肠中的L.motocytogenes.%An oscillatory-flow PCR microfluidics has been sucessfully developed for rapid detection of foodborne bacterial pathogens ( Listeria monocytogene).The device mainly coasists of the LabView-based temperature control and measurement system, three pieces of copper block and polytetrafluoroethylene (PTFE) capillary.On this device, three copper blocks provided the temperatures for PCR, and the PTFE capillary was embedded in three copper blocks as the PCR reaction channel.DNA amplifications were performed within individual capillaries when the PCR solutions were driven by the precision syringe pump in an oscillatory way.Successful amplification of 135 bp DNA fragment was achieved in less than 14 min when the flow rate of 300 μL/min was used, and the detection limit of Listeria monocytogene ( L.monocytogenes) was 10 cuf/mL.This system has great potential to perform the high-throughput PCR amplification in separate capillaries.As a demonstration, simultaneous detection of L.rnonocytogenes in water, milk, banana and sausage was performed on the developed oscillatory-flow PCR microfluidics.