期刊
TALANTA
卷 122, 期 -, 页码 246-250出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2014.01.053
关键词
Continuous-flow PCR; 16S rDNA; Hybridization
资金
- NSFC [21377026, 20975025]
- Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China [708031]
Rapid identification of clinical pathogens is the initial and essential step for antimicrobial therapy. Herein, we successfully developed a microfluidic device which combines high-throughput continuousflow PCR and DNA hybridization for the detection of various bacterial pathogens. Universal primers were designed based on the conserved regions of bacterial 16S ribosomal DNA (16S rDNA), and specific probes were designed from a variable region of 16S rDNA within the amplicon sequences. In the chip operation, after the continuous flow PCR was achieved in the first microfluidic chip, the product was directly introduced into a hybridization chip integrated with microarray containing the immobilized DNA probes. The target-probe hybridization was completed within 1 h at 55 degrees C, and fluorescence signals were obtained as the readout. The presented device is simple, versatile and with less sample consumption compared with traditional instruments. It can perform high-throughput bacteria detections continuously in a single assay, which makes it a promising platform for clinical bacteria identifications. (c) 2014 Elsevier B.V. All rights reserved.
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