An ultrasensitive and rapid "sample-to-answer" microsystem for on-site monitoring of SARS-CoV-2 in aerosols using "in situ" tetra-primer recombinase polymerase amplification.

Airborne transmissibility of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has highlighted the urgent need for aerosol monitoring of SARS-CoV-2 to prevent sporadic outbreaks of COVID-19. The inadequate sensitivity of conventional methods and the lack of an on-site detection system limited the practical SARS-CoV-2 monitoring of aerosols in public spaces. We have developed a novel SARS-CoV-2-in-aerosol monitoring system (SIAMs) which consists of multiple portable cyclone samplers for collecting aerosols from several venues and a sensitive "sample-to-answer" microsystem employing an integrated cartridge for the analysis of SARS-CoV-2 in aerosols (iCASA) near the sampling site. By seamlessly combining viral RNA extraction based on a chitosan-modified quartz filter and "in situ" tetra-primer recombinase polymerase amplification (tpRPA) into an integrated microfluidic cartridge, iCASA can provide an ultra-high sensitivity of 20 copies/mL, which is nearly one order of magnitude greater than that of the commercial kit, and a short turnaround time of 25 min. By testing various clinical samples of nasopharyngeal swabs, saliva, and exhaled breath condensates obtained from 23 COVID-19 patients, we demonstrate that the positive rate of our system was 3.3 times higher than those of the conventional method. Combining with multiple portable cyclone samplers, we detected 52.2% (12/23) of the aerosol samples, six times higher than that of the commercial kit, collected from the isolation wards of COVID-19 patients, demonstrating the excellent performance of our system for SARS-CoV-2-in-aerosol monitoring. We envision the broad application of our microsystem in aerosol monitoring for fighting the COVID-19 pandemic.

Emerging membrane technologies and disinfection methods for efficient removal of waterborne pathogens during the COVID-19 pandemic and post-pandemic

Viruses and other microorganisms can enter water sources from different routes and cause pollution and irreparable damage. So, cost-effective and efficient systems for providing safe water are necessary. Efficient filtration systems based on antimicrobial materials have received a lot of attention in this regard. A wide range of materials play an important role in the production of efficient water filtration systems. Metal and metal oxide particles with anti-viral and antimicrobial properties comprising Cu, Cu2O, Ag, TiO2, and ZnO play a valuable role in the preparation of water filtration systems. Biopolymers such as cellulose or carbon nanomaterials like graphene or its derivatives have been reported to provide safe water. In this review, we summarize the use of diverse materials in the preparation of efficient filtration-based systems like membranes and paper filters for water treatment. Pathogen-containing water samples were effectively disinfected using the prepared water disinfection systems.

All-solid-state paper-based potentiometric combined sensor modified with reduced graphene oxide (rGO) and molecularly imprinted polymer for monitoring losartan drug in pharmaceuticals and biological samples.

A cost-effective, highly selective and sensitive paper-based potentiometric combined sensor for losartan potassium drug (LOS) is fabricated, characterized and used for the drug monitoring. The sensor consists of 2 strips of filter paper (20 × 5 mm each) as platform, each imprinted with 4 mm diameter circular spot of carbon. One carbon spot is covered by a reduced graphene oxide (rGO) for use as a substrate for the recognition sensor and the other without rGO is used for the reference electrode. LOS molecularly imprinted drug polymer (MIP) is applied onto the graphene oxide containing strip to act as a drug recognition sensing material and a solid-state polyvinyl butyral (PVB) is applied onto the second carbon spot to act as a reference electrode. Performance characteristics of the combined sensor are examined with chronopotentiometry (CP) and electrochemical impedance spectroscopy (EIS). Increase effect of rGO on the interfacial double-layer capacitance of the sensing membrane and consequently on the potential stability is confirmed. The developed combined sensor (strip cell) displays a Nernstian slope of -58.2 ± 0.3 mV/decade (R2 = 0.9994) over the linear range 8.5 × 10-7 - 6.9 × 10-2 M with a detection limit of 2.7 ± 0.3 × 10-7 M. The sensor shows remarkable selectivity toward various related compounds especially those commonly used by the COVID-19 patients such as paracetamol, ascorbic acid and dextromethorphan. The assay method is validated and proved to be satisfactory for direct potentiometric determination of LOS-K in some pharmaceutical formulations and in spiked human urine samples. An average recovery of 96.3 ± 0.3-98.7 ± 0.6% of the nominal or spiked concentration and a mean relative standard deviation of ±0.6% are obtained. The use of an indicating and a reference electrodes combined into a single flexible disposable paper platform enables applications to a minimum sample volume due to the close proximity of the responsive membrane and the liquid junction. The efficiency of the proposed sensor in complex urine matrix suggests its application in hospitals for rapid diagnosis of overdose patients and for quality control/quality assurance tests in pharmaceutical industry.

Evaluation of filter paper to transport oro/nasopharyngeal samples to detect SARS-CoV-2 by RT-qPCR.

PURPOSE: To evaluate filter paper as a means to transport oro/nasopharyngeal samples from laboratories with few resources for SARS-CoV-2 detection by RT-qPCR in a central laboratory that usually performs this technique as routine. METHODS: A total of 40 specimens were evaluated in parallel by RT-qPCR carried out after RNA extraction using two different protocols: direct RNA extraction (Protocol A - reference method) and RNA extraction after impregnation in filter paper (Protocol B). RESULTS: The RT-qPCR for SARS-CoV-2 using Protocol B presented 97.22% (35/36) of agreement for SARS-CoV-2-positive samples when compared to the reference method (Protocol A), even for specimens with low viral load (increased Ct values). Noteworthy, three clinical specimens which were categorized as inconclusive by Protocol A presented amplification of both N1 and N2 targets using Protocol B, presenting positive results for SARS-CoV-2. CONCLUSION: The use of filter paper to transport oro/nasopharyngeal clinical samples presented very satisfactory results to detect SARS-CoV-2 by RT-qPCR. In addition, it proved to be a feasible and sensitive approach, being able to generate the detection of SARS-CoV-2 even at low concentrations, without presenting false-negative results.

A dried blood spot protocol for high throughput analysis of SARS-CoV-2 serology based on the Roche Elecsys anti-N assay.

BACKGROUND: Since 2020 SARS-CoV-2 spreads pandemically, infecting more than 119 million people, causing >2·6 million fatalities. Symptoms of SARS-CoV-2 infection vary greatly, ranging from asymptomatic to fatal. Different populations react differently to the disease, making it very hard to track the spread of the infection in a population. Measuring specific anti-SARS-CoV-2 antibodies is an important tool to assess the spread of the infection or successful vaccinations. To achieve sufficient sample numbers, alternatives to venous blood sampling are needed not requiring medical personnel or cold-chains. Dried-blood-spots (DBS) on filter-cards have been used for different studies, but not routinely for serology. METHODS: We developed a semi-automated protocol using self-sampled DBS for SARS-CoV-2 serology. It was validated in a cohort of matched DBS and venous-blood samples (n = 1710). Feasibility is demonstrated with two large serosurveys with 10247 company employees and a population cohort of 4465 participants. FINDINGS: Sensitivity and specificity reached 99·20% and 98·65%, respectively. Providing written instructions and video tutorials, 99·87% (4465/4471) of the unsupervised home sampling DBS cards could be analysed. INTERPRETATION: DBS-sampling is a valid and highly reliable tool for large scale serosurveys. We demonstrate feasibility and accuracy with a large validation cohort including unsupervised home sampling. This protocol might be of big importance for surveillance in resource-limited settings, providing low-cost highly accurate serology data. FUNDING: Provided by Bavarian State Ministry of Science and the Arts, LMU University-Hospital; Helmholtz-Centre-Munich, German Ministry for Education and Research (project01KI20271); University of Bonn; University of Bielefeld; the Medical Biodefense Research Program of Bundeswehr-Medical-Service; Euroimmun, RocheDiagnostics provided discounted kits and machines.

The use of exhaled nitric oxide and peak expiratory flow to demonstrate improved breathability and antimicrobial properties of novel face mask made with sustainable filter paper and Folium Plectranthii amboinicii oil: additional option for mask shortage during COVID-19 pandemic.

BACKGROUND: Medical face masks are integral personal protective equipment against infectious airborne disease and become scarce during epidemic outbreaks such as COVID-19. A novel, sustainably manufactured face mask with antimicrobial and anti-inflammatory properties from oil of Folium Plectranthii amboinicii can be an effective alternative to internationally sold masks. METHODS: This prospective, randomized study assigned subjects (n=67) to either conventional surgical face mask or Lamdong Medical College (LMC) face mask for three hours. Fractional concentration of nitric oxide in exhaled breath (FENO) and peak expiratory flow (PEF) was measured before and after mask use. Subjective reporting on respiratory symptoms was also analyzed. Masks were then incubated and analyzed for microorganism growth. RESULTS: Subjects assigned the LMC mask had a lowered FENO (p<0.05) compared to conventional face masks after mask wearing. Subjects with LMC mask use reported higher comfortability (p<0.05), breathability (p<0.05), and lower allergy symptoms (p<0.05). The LMC mask has visually less microorganism growth in the cultured medium, measured by sterile ring radius. CONCLUSIONS: The LMC face mask is a renewably manufactured personal protective tool with antibacterial capacity that can serve as an effective alternative to internationally sold surgical face mask during shortage of mask due to COVID-19.

Validation of dried blood spot sample modifications to two commercially available COVID-19 IgG antibody immunoassays.

Aim: Coronavirus disease 2019 antibody testing often relies on venous blood collection, which is labor-intensive, inconvenient and expensive compared with finger-stick capillary dried blood spot (DBS) collection. The purpose of our work was to determine if two commercially available anti-severe acute respiratory syndrome coronavirus 2 enzyme-linked immunosorbent assays for IgG antibodies against spike S1 subunit and nucleocapsid proteins could be validated for use with DBS. Materials & methods: Kit supplied reagents were used to extract DBS, and in-house DBS calibrators were included on every run. Results: Positive/negative concordance between DBS and serum was 100/99.3% for the spike S1 subunit assay and 100/98% for the nucleocapsid assay. Conclusion: Validation of the DBS Coronavirus disease 2019 IgG antibody assays demonstrated that serum and DBS can produce equivalent results with minimal kit modifications.