Real-time PCR detection of Listeria monocytogenes using an integrated microfluidics platform
2004; Elsevier BV; Volume: 107; Issue: 1 Linguagem: Inglês
10.1016/j.snb.2004.10.022
ISSN1873-3077
AutoresNathaniel C. Cady, Scott Stelick, Madanagopal V. Kunnavakkam, Carl A. Batt,
Tópico(s)Microfluidic and Capillary Electrophoresis Applications
ResumoA miniaturized, fully automated, PCR-based detection system has been developed for the rapid detection of bacterial pathogens. Monolithic DNA purification/real-time PCR silicon chips were fabricated and tested for their ability to purify and detect the pathogeneic bacterium Listeria monocytogenes. Using silica-coated microstructures, nucleic acids could be selectively bound, washed and eluted for subsequent real-time PCR. These microstructures were included in an integrated detection microchip containing two distinct regions, one for DNA purification and one for real-time PCR. Using an automated detection platform with integrated microprocessor, pumps, valves, thermocycler and fluorescence detection modules, microchips were used to purify and detect bacterial DNA by real-time PCR amplification using SYBR Green fluorescent dye. Between 104 and 107 L. monocytogenes cells could be detected using this system with an average turnaround time of 45 min.
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