We report an electrochemical biosensor combined with recombinase polymerase amplification (RPA) for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2. The electrochemical biosensor based on a multi-microelectrode array allows the detection of multiple target genes by differential pulse voltammetry. The RPA reaction involves hybridization of the RPA amplicon with thiol-modified primers immobilized on the working electrodes, which leads to a reduction of current density as amplicons accumulate. The assay results in shorter “sample-to-answer” times than conventional PCR without expensive thermo-cycling equipment. The limits of detection are about 0.972 fg/μL (RdRP gene) and 3.925 fg/μL (N gene), which are slightly lower than or comparable to that of RPA assay results obtained by gel electrophoresis without post-amplification purification. The combination of electrochemical biosensors and the RPA assay is a rapid, sensitive, and convenient platform that can be potentially used as a point-of-care test for the diagnosis of COVID-19.
A surface-enhanced
Raman scattering-based lateral flow assay (SERS-LFA)
technique has been developed for the rapid and accurate diagnosis
of scrub typhus. Lateral flow kits for the detection of O.
tsutsugamushi IgG (scrub typhus biomarker) were fabricated,
and the calibration curve for various standard clinical sera concentrations
were obtained by Raman measurements. The clinical sera titer values
were determined by fitting the Raman data to the calibration curve.
To assess the clinical feasibility of the proposed method, SERS-LFA
assays were performed on 40 clinical samples. The results showed good
agreement with those of the standard indirect immunofluorescence assay
(IFA) method. SERS-LFA has many advantages over IFA including the
less sample volume, simpler assay steps, shorter assay time, more
systematic quantitative analysis, and longer assay lifetime. As SERS
strips can be easily integrated with a miniaturized Raman spectrophotometer,
field serodiagnosis is also more feasible.
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