In this pilot study, we characterize and evaluate 3D-printed swabs for the collection of nasopharyngeal and oropharyngeal secretion samples for the SARS-CoV-2 detection. Swabs are made with the fused deposition modeling technique using the biopolymer polylactic acid (PLA) which is a medical-grade, biodegradable and low-cost material. We evaluated six swabs with mechanical tests in a laboratory and in an Adult Human Simulator performed by healthcare professionals. We proved the adequacy of the PLA swab to be used in the gold standard reverse transcriptase-polymerase chain reaction (qRT-PCR) for viral RNA detection. Then, we did in vitro validation for cell collection using the 3D-printed swabs and RNA extraction for samples from 10 healthy volunteers. The 3D-printed swabs showed good flexibility and maneuverability for sampling and at the same time robustness to pass into the posterior nasopharynx. The PLA did not interfere with the RNA extraction process and qRT-PCR test. When we evaluated the expression of the reference gene (RNase P) used in the SARS-CoV-2 detection, the 3D-printed swabs showed good reproducibility in the threshold cycle values (Ct = 23.5, range 19–26) that is comparable to control swabs (Ct = 24.7, range 20.8–32.6) with p value = 0.47. The 3D-printed swabs demonstrated to be a reliable, and an economical alternative for mass use in the detection of SARS-CoV-2.
In this work, a microfluidic device with a three-electrode system is used to measure the electrical impedance spectrum of micron-sized particles suspended in Phosphate Buffered Saline (PBS). We use polymer particles of 15 μm and 48 μm of diameter, as they can mimic biological cells in suspension. The main purpose is to study the influence of the particle concentration on impedance parameters and to establish the basis for biological cell suspensions assays. An electric equivalent circuit model is used to describe the experimental impedance data. Results show a high dependence of electrical parameters with the mass fraction and particle size. The model considers the resistance of the bulk solution and the electrical double layer, as the two essential electrical elements. Results show a maximum error of 3% by fitting the measurements to the proposed model. This proposal could be of interest for cytotoxicity assays.
Analyzing the electrical double layer (EDL) in electrical impedance spectroscopy (EIS) measurement at low frequencies remains a challenging task for sensing purposes. In this work, we propose two approaches to deal with the EDL in measuring impedance for particles and non-adherent cells in an electrolytic suspension. The first approach is a simple procedure to compute a normalized electrical impedance spectrum named dispersed medium index (DMi). The second is the EIS modeling through an equivalent electric circuit based on the so-called effective capacitance (Cef), which unifies the EDL phenomena. Firstly, as an experiment under controlled conditions, we examine polymer particles of 6, 15, and 48 μm in diameter suspended in a 0.9% sodium chloride solution. Subsequently, we used K-562 cells and leukocytes suspended in a culture medium (RPMI-1640 supplemented) for a biological assay. As the main result, the DMi is a function of the particle concentration. In addition, it shows a tendency with the particle size; regardless, it is limited to a volume fraction of 0.03 × 10−4 to 58 × 10−4. The DMi is not significantly different between K-562 cells and leukocytes for most concentrations. On the other hand, the Cef exhibits high applicability to retrieve a function that describes the concentration for each particle size, the K-562 cells, and leukocytes. The Cef also shows a tendency with the particle size without limitation within the range tested, and it allows distinction between the K-562 and leukocytes in the 25 cells/µL to 400 cells/µL range. We achieved a simple method for determining an Cef by unifying the parameters of an equivalent electrical circuit from data obtained with a conventional potentiostat. This simple approach is affordable for characterizing the population of non-adherent cells suspended in a cell culture medium.
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