Microfluidic chip integrating high throughput continuous-flow PCR and DNA hybridization for bacteria analysis.
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 continuous-flow 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 1h at 55 °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.
Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention (LAP(3)), Department of Environmental Science & Engineering, Institute of Biomedical Science, Fudan University, Shanghai, 200433, PR China.
Xiran
X
Jiang
Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), School of Biomedical Engineering, State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiao Tong University, Shanghai, 200240, China.
Ning
N
Shao
Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention (LAP(3)), Department of Environmental Science & Engineering, Institute of Biomedical Science, Fudan University, Shanghai, 200433, PR China; Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, PR China.
Wenwen
W
Jing
Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), School of Biomedical Engineering, State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiao Tong University, Shanghai, 200240, China.
Shengce
S
Tao
Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention (LAP(3)), Department of Environmental Science & Engineering, Institute of Biomedical Science, Fudan University, Shanghai, 200433, PR China.
Sixiu
S
Liu
Shanghai Key Laboratory of Atmospheric Particle Pollution and Prevention (LAP(3)), Department of Environmental Science & Engineering, Institute of Biomedical Science, Fudan University, Shanghai, 200433, PR China. Electronic address: gsui@fudan.edu.cn.
Guodong
G
Sui
D015342:DNA Probes / Q000737:chemistry*; D004269:DNA, Bacterial / Q000032:analysis*; D057166:High-Throughput Screening Assays / Q000379:methods; D046210:Microfluidic Analytical Techniques / Q000379:methods; D044085:Microfluidics / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction
D015342:DNA Probes; D004269:DNA, Bacterial
16S rDNA; Continuous-flow PCR; Hybridization