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1.
Sensors (Basel) ; 19(16)2019 Aug 14.
Article in English | MEDLINE | ID: mdl-31416185

ABSTRACT

Pre-treatment screening of individuals for human leukocyte antigens (HLA) HLA-B*57:01 is recommended for the prevention of life-threatening hypersensitivity reactions to abacavir, a drug widely prescribed for HIV treatment. However, the implementation of screening in clinical practice is hindered by the slow turnaround time and high cost of conventional HLA genotyping methods. We have developed a biosensor platform using interdigitated electrode (IDE) functionalized with a monoclonal antibody to detect cells expressing HLA-B*57:01. This platform was evaluated using cell lines and peripheral blood mononuclear cells expressing different HLA-B alleles. The functionalized IDE sensor was able to specifically capture HLA-B*57:01 cells, resulting in a significant change in the impedance magnitude in 20 min. This IDE platform has the potential to be further developed to enable point-of-care HLA-B*57:01 screening.


Subject(s)
Biosensing Techniques/methods , Dideoxynucleosides/adverse effects , Drug Hypersensitivity/prevention & control , HLA-B Antigens/analysis , Leukocytes, Mononuclear/metabolism , Alleles , Antibodies, Immobilized/chemistry , Antibodies, Immobilized/immunology , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/immunology , Dideoxynucleosides/therapeutic use , Drug Hypersensitivity/etiology , Electrochemical Techniques , Electrodes , HIV Infections/drug therapy , HLA-B Antigens/genetics , HLA-B Antigens/immunology , Humans
2.
Sci Rep ; 5: 16485, 2015 Nov 09.
Article in English | MEDLINE | ID: mdl-26548806

ABSTRACT

The application of microfluidic devices in diagnostic systems is well-established in contemporary research. Large specific surface area of microspheres, on the other hand, has secured an important position for their use in bioanalytical assays. Herein, we report a combination of microspheres and microfluidic disk in a unique hybrid platform for highly sensitive and selective detection of dengue virus. Surface engineered polymethacrylate microspheres with carefully designed functional groups facilitate biorecognition in a multitude manner. In order to maximize the utility of the microspheres' specific surface area in biomolecular interaction, the microfluidic disk was equipped with a micromixing system. The mixing mechanism (microballoon mixing) enhances the number of molecular encounters between spheres and target analyte by accessing the entire sample volume more effectively, which subsequently results in signal amplification. Significant reduction of incubation time along with considerable lower detection limits were the prime motivations for the integration of microspheres inside the microfluidic disk. Lengthy incubations of routine analytical assays were reduced from 2 hours to 5 minutes while developed system successfully detected a few units of dengue virus. Obtained results make this hybrid microsphere-microfluidic approach to dengue detection a promising avenue for early detection of this fatal illness.


Subject(s)
Dengue Virus , Dengue/diagnosis , Microfluidics/methods , Microspheres , Enzyme-Linked Immunosorbent Assay/methods , Humans , Methacrylates/chemistry , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods , Microfluidics/instrumentation , Particle Size , Polymethacrylic Acids/chemistry , Reproducibility of Results , Sensitivity and Specificity , Time Factors
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