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Micromachines (Basel) ; 12(10)2021 Sep 24.
Article in English | MEDLINE | ID: covidwho-1438668


This paper reports the design, development, and testing of a novel, yet simple and low-cost portable device for the rapid detection of SARS-CoV-2. The device performs loop mediated isothermal amplification (LAMP) and provides visually distinguishable images of the fluorescence emitted from the samples. The device utilises an aluminium block embedded with a cartridge heater for isothermal heating of the sample and a single-board computer and camera for fluorescence detection. The device demonstrates promising results within 20 min using clinically relevant starting concentrations of the synthetic template. Time-to-signal data for this device are considerably lower compared to standard quantitative Polymerase Chain Reaction(qPCR) machine (~10-20 min vs. >38 min) for 1 × 102 starting template copy number. The device in its fully optimized and characterized state can potentially be used as simple to operate, rapid, sensitive, and inexpensive platform for population screening as well as point-of-need severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) detection and patient management.

Analyst ; 145(23): 7680-7686, 2020 Nov 23.
Article in English | MEDLINE | ID: covidwho-798256


This work reports the development of a rapid, simple and inexpensive colorimetric paper-based assay for the detection of the severe acute respiratory symptom coronavirus 2 (SARS-CoV-2) humanized antibody. The paper device was prepared with lamination for easy sample handling and coated with the recombinant SARS-CoV-2 nucleocapsid antigen. This assay employed a colorimetric reaction, which is followed by horseradish peroxidase (HRP) conjugated detecting antibody in the presence of the 3,3',5,5'-tetramethylbenzidine (TMB) substrate. The colorimetric readout was evaluated and quantified for specificity and sensitivity. The characterization of this assay includes determining the linear regression curve, the limit of detection (LOD), the repeatability, and testing complex biological samples. We found that the LOD of the assay was 9.00 ng µL-1 (0.112 IU mL-1). The relative standard deviation was approximately 10% for a sample number of n = 3. We believe that our proof-of-concept assay has the potential to be developed for clinical screening of the SARS-CoV-2 humanized antibody as a tool to confirm infected active cases or to confirm SARS-CoV-2 immune cases during the process of vaccine development.

Antibodies, Monoclonal, Humanized/blood , Antibodies, Viral/blood , COVID-19 Testing/methods , Colorimetry/methods , Enzyme-Linked Immunosorbent Assay/methods , Paper , SARS-CoV-2/immunology , Antibodies, Monoclonal, Humanized/immunology , Antibodies, Viral/immunology , Armoracia/enzymology , Benzidines/chemistry , COVID-19/diagnosis , COVID-19 Testing/instrumentation , Colorimetry/instrumentation , Coronavirus Nucleocapsid Proteins/immunology , Enzyme-Linked Immunosorbent Assay/instrumentation , Horseradish Peroxidase/chemistry , Humans , Limit of Detection , Phosphoproteins/immunology , Proof of Concept Study , SARS-CoV-2/chemistry