Multichannel oscillatory-flow multiplex PCR microfluidics for high-throughput and fast detection of foodborne bacterial pathogens

Content Provider	Semantic Scholar
Author	Zhang, Chunsun Wang, Haiying Xing, Da
Copyright Year	2011
Abstract	In the field of continuous-flow PCR, the amplification throughput in a single reaction solution is low and the single-plex PCR is often used. In this work, we reported a flow-based multiplex PCR microfluidic system capable of performing high-throughput and fast DNA amplification for detection of foodborne bacterial pathogens. As a demonstration, the mixture of DNA targets associated with three different foodborne pathogens was included in a single PCR solution. Then, the solution flowed through microchannels incorporated onto three temperature zones in an oscillatory manner. The effect factors of this oscillatory-flow multiplex PCR thermocycling have been demonstrated, including effects of polymerase concentration, cycling times, number of cycles, and DNA template concentration. The experimental results have shown that the oscillatory-flow multiplex PCR, with a volume of only 5 μl, could be completed in about 13 min after 35 cycles (25 cycles) at 100 μl/min (70 μl/min), which is about one-sixth of the time required on the conventional machine (70 min). By using the presently designed DNA sample model, the minimum target concentration that could be detected at 30 μl/min was 9.8 × 10−2 ng/μl (278-bp, S. enterica), 11.2 × 10−2 ng/μl (168-bp, E. coli O157: H7), and 2.88 × 10−2 ng/μl (106-bp, L. monocytogenes), which corresponds to approximately 3.72 × 104 copies/μl, 3.58 × 104 copies/μl, and 1.79 × 104 copies/μl, respectively. This level of speed and sensitivity is comparable to that achievable in most other continuous-flow PCR systems. In addition, the four individual channels were used to achieve multi-target PCR analysis of three different DNA samples from different food sources in parallel, thereby achieving another level of multiplexing.
Starting Page	885
Ending Page	897
Page Count	13
File Format	PDF HTM / HTML
DOI	10.1007/s10544-011-9558-y
PubMed reference number	21691814
Journal	Medline
Volume Number	13
Alternate Webpage(s)	http://laser.scnu.edu.cn/xingdaPDF/2011pdf/Zhang%20Chunsun%20Biomed%20Microdevices%202011.pdf
Journal	Biomedical microdevices
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article