RESUMEN
In this study, a rapid diagnosis platform was developed for the detection of Escherichia coli O157:H7. An electrical double layer (EDL)-gated field-effect transistor-based biosensor (BioFET) as a point-of-care testing device is demonstrated with its high sensitivity, portability, high selectivity, quick response, and ease of use. The specially designed ssDNA probe was immobilized on the extended gate electrode to bind the target complementary DNA segment of E. coli, resulting in a sharp drain current change within minutes. The limit of detection for target DNA is validated to a concentration of 1 fM in buffer solution and serum. Meanwhile, the results of a Kelvin probe force microscope were shown to have reduced surface potential of the DNA immobilized sensors before and after the cDNA detection, which is consistent with the decreased drain current of the BioFET. A 1.2 kb E. coli duplex DNA synthesized in plasmid was sonicated and detected in serum samples with the sensor array. Gel electrophoresis was used to confirm the efficiency of sonication by elucidating the length of DNA. Those results show that the EDL-gated BioFET system is a promising platform for rapid identification of pathogens for future clinical needs.
Asunto(s)
Técnicas Biosensibles , Infecciones por Escherichia coli , Escherichia coli O157 , Humanos , Técnicas Biosensibles/métodos , ADN de Cadena Simple , Electrodos , Escherichia coli O157/genética , ADN Bacteriano/genéticaRESUMEN
In light of the swift outspread and considerable mortality, coronavirus disease 2019 (COVID-19) necessitates a rapid screening tool and a precise diagnosis. Saliva is considered as an alternative specimen to detect the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) since the viral load is comparable to what are found in a throat and a nasal cavity. The electrical double layer (EDL)-gated field-effect transistor-based biosensor (BioFET) emerges as a promising candidate for salivary COVID-19 tests due to a high sensitivity, a portable configuration, a label-free operation, and a matrix insensitivity. In this work, the authors utilize EDL-gated BioFETs to detect complementary DNAs (cDNAs) and viral RNAs with various testing conditions such as switches of probes, temperature treatments, and matrices. The selectivity is confirmed with cDNA and noncomplementary DNA (ncDNA), exhibiting an eightfold difference in electrical signals. The matrix insensitivity is evaluated, and BioFETs successfully validate the detection of SARS-CoV-2 N-gene RNA down to 1 fm in diluted human saliva with a 95°C- and a 25°C-treatment, respectively. This proposed system has a high potential to be deployed for an on-site COVID-19 screening, improving the disease control and benefitting frontline healthcare system.
RESUMEN
In this experimental study, a portable biosensor was developed to detect ß-human chorionic gonadotropin (ß-hCG), which is extensively used in pregnancy tests and serves as a biomarker for ectopic pregnancy. The sensor used is an electric-double-layer field-effect transistor biosensor with the extended-gate design. Bias voltage is applied on the sensor to measure the resulting drain current signals. Gold electrode surface is functionally activated with an anti-ß-hCG antibody to capture ß-hCG protein. Fluorescence imaging technique is utilized to confirm the surface functionalization. The biosensor demonstrates a dynamically wide range of molecules as detection targets at very low sample concentrations, which shows the potential to detect ectopic pregnancy in very early stages and easily keep track of its periodic changes. It can be produced en masse and does not use additional labels/reagents or pre-processing techniques for the sample. This biosensor can significantly reduce the manufacturing costs and is comparable with the currently available commercial ß-hCG assays. It is suitable for early diagnosis of ectopic pregnancy with low cost and easy operation at home with urine samples.