We have developed a novel multiplexed bead-based mesofluidic system (MBMS) based on the specific recognition events on the surface of a series of microbeads (diameter 250 μm) arranged in polydimethylsiloxane (PDMS) microchannels (diameter 300 μm) with the predetermined order and assembled an apparatus implementing automatically the high-throughput bead-based assay and further demonstrated its feasibility and flexibility of gene diagnosis and genotyping, such as β-thalassemia mutation detection and HLA-DQA genotyping. The apparatus, consisting of bead-based mesofluidic PDMS chip, liquid-processing module, and fluorescence detection module, can integrate the procedure of automated-sampling, hybridization reactions, washing, and in situ fluorescence detection. The results revealed that MBMS is fast, has high sensitivity, and can be automated to carry out parallel and multiplexed genotyping and has the potential to open up new routes to flexible, high-throughput approaches for bioanalysis.
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Multiplexed bead-based mesofluidic system for gene diagnosis and genotyping.
Anal Chem
December 2010
Lab of Biosystems and Microanalysis, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai, 200237, China.
We have developed a novel multiplexed bead-based mesofluidic system (MBMS) based on the specific recognition events on the surface of a series of microbeads (diameter 250 μm) arranged in polydimethylsiloxane (PDMS) microchannels (diameter 300 μm) with the predetermined order and assembled an apparatus implementing automatically the high-throughput bead-based assay and further demonstrated its feasibility and flexibility of gene diagnosis and genotyping, such as β-thalassemia mutation detection and HLA-DQA genotyping. The apparatus, consisting of bead-based mesofluidic PDMS chip, liquid-processing module, and fluorescence detection module, can integrate the procedure of automated-sampling, hybridization reactions, washing, and in situ fluorescence detection. The results revealed that MBMS is fast, has high sensitivity, and can be automated to carry out parallel and multiplexed genotyping and has the potential to open up new routes to flexible, high-throughput approaches for bioanalysis.
View Article and Find Full Text PDFMultiplexed bead-based mesofluidic system for detection of food-borne pathogenic bacteria.
Appl Environ Microbiol
November 2009
Lab of Biosystems and Microanalysis, East China University of Science & Technology, Shanghai 200237, China.
In the present study, a simple and rapid multiplexed bead-based mesofluidic system (BMS) was developed for simultaneous detection of food-borne pathogenic bacteria, including Staphylococcus aureus, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella, Enterobacter sakazakii, Shigella, Escherichia coli O157:H7, and Campylobacter jejuni. This system is based on utilization of isothiocyanate-modified microbeads that are 250 mum in diameter, which were immobilized with specific amino-modified oligonucleotide probes and placed in polydimethylsiloxane microchannels. PCR products from the pathogens studied were pumped into microchannels to hybridize with the oligonucleotide-modified beads, and hybridization signals were detected using a conventional microarray scanner.
View Article and Find Full Text PDFBead-based mesofluidic system for residue analysis of chloramphenicol.
J Agric Food Chem
November 2008
Laboratory of Biosystem and Microanalysis, Department of Food Engineering and Science, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China.
A highly selective and sensitive mesofluidic immunoassay system based on competitive immunoassay in polydimethylsiloxane (PDMS) channels was developed. This immunoassay system was successfully applied to quantificationally detect chloramphenicol (CAP) in animal foods. The glass beads (Ø 250 microm) were amino-silane modified, covalently precoated with chloramphenicol succinate, and then infused into the microchannels (Ø 300 microm); the CAP molecules of samples or standards in flow solution competed for CAP antibody with the CAP immobilized on the beads.
View Article and Find Full Text PDFDifferent strategies of covalent attachment of oligonucleotide probe onto glass beads and the hybridization properties.
Appl Biochem Biotechnol
January 2009
Laboratory of Bioanalysis and Biosystems, State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, China.
The glass bead is a new biochip support material for immobilization biomolecules, due to its independence and convenient rearrangement. In order to optimize the immobilization efficiency of oligonucleotides onto glass beads and obtain the highest hybridization efficiency, three commonly used coupling strategies have been studied for covalently attaching oligonucleotides onto large glass beads. Glass beads with 250 microm diameter were amino-silaned with 2% 3-aminopropyltrimethoxysilane (APTMS) and then reacted separately with glutaraldehyde, succinic anhydride and 1,4-phenylene diisothiocyanate (PDITC) to derive CHO beads, COOH beads and isothiocyanate-modified beads (NCS-Beads) accordingly.
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