Severe acute respiratory syndrome coronavirus 2 detection by real time polymerase chain reaction using pooling strategy of nasal samples.

COVID-19 is a life-threatening multisistemic infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Infection control relies on timely identification and isolation of infected people who can alberg the virus for up to 14 days, providing important opportunities for undetected transmission. This note describes the application of rRT-PCR test for simpler, faster and less invasive monitoring of SARS-CoV-2 infection using pooling strategy of samples. Seventeen positive patients were provided with sterile dry swabs and asked to self-collected 2 nasal specimens (#NS1 and #NS2). The #NS1 was individually placed in a single tube and the #NS2 was placed in another tube together with 19 NSs collected from 19 negative patients. Both tubes were then tested with conventional molecular rRT-PCR and the strength of pooling nasal testing was compared with the molecular test performed on the single NS of each positive patient. The pooling strategy detected SARS-CoV-2 RNA to a similar extent to the single test, even when Ct value is on average high (Ct 37-38), confirming that test sensibility is not substantially affected even if the pool contains only one low viral load positive sample. Furthermore, the pooling strategy have benefits for SARS-CoV-2 routinary monitoring of groups in regions with a low SARS-CoV-2 prevalence.

A simple pooling salivary test for SARS-CoV-2 diagnosis: A Columbus' egg?

Saliva is an appropriate specimen for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) diagnosis. The possibility of pooling samples of saliva, using non-invasive bibula strips for sampling, was explored employing Bovine coronavirus (BCoV) spiked saliva. In laboratory, up to 30 saliva-soaked strips were pooled in a single tube with 2 mL of medium. After quick adsorption with the medium and vortexing, the liquid was collected and tested with a quantitative molecular assay to quantify viral RNA genome copies. On testing of single and pooled strips, the difference between the median threshold cycles (Ct) value of test performed on the single positive saliva sample and the median Ct value obtained on the pool of 30 strips, was 3.21 cycles. Saliva pooling with bibula strips could allow monitoring of COVID-19 on a large scale, reducing costs for the health bodies in terms of medical material and skilled personnel. Finally, saliva sampling is noninvasive and less traumatic than nasopharyngeal swabs and can be self-collected.

Evaluation of virucidal activity of fabrics using feline coronavirus.

Severe Acute Respiratory Syndrome Coronavirus type 2 (SARS-CoV-2) is an enveloped RNA virus responsible for the 2019 coronavirus disease (COVID-19) that represents a global health threat, causing an ongoing pandemic in many countries and territories. WHO recommendations emphasize the importance of all personal protective equipment (PPE) that can interrupt COVID-19 transmission. The textile industry and scientists are developing hygienic fabrics by the addition of or treatment with various antimicrobial and antiviral compounds. Methods for determining the antiviral activity of fabrics are reported in the International Standards Organization (ISO) 18184 (2019) guidelines. Three different fabric samples treated with silver derivate, copper derivative and a not treated cotton fabric used as control were examined and put in contact with a suspension of feline coronavirus (FCoV). After 2 h of incubation a significant decrease of viral titer, as high as 3.25 log10 Tissue Culture Infectious Dose (TCID)50/50 µl, in feline cells was observed in treated fabrics, with respect to not treated fabrics. In this study, we optimized laboratory methods to evaluate the virucidal activity of silver- and copper treated cotton- based fabrics against coronavirus, using FCoV suitable as a surrogate of SARS-CoV-2 but safe for laboratory technicians.

Virucidal and antiviral effects of Thymus vulgaris essential oil on feline coronavirus.

Feline infectious peritonitis (FIP) is a fatal systemic disease of felids caused by a Coronavirus (CoV) (FIPV). In spite of its clinical relevance and impact on feline health, currently the therapeutic possibilities for treatment of FIP in cats are limited. The emergence of the pandemic Severe Respiratory Syndrome (SARS) coronavirus (CoV) type 2 (SARS-CoV-2), etiological agent of the 2019 Coronavirus Disease (COVID-19), able to infect a broad spectrum of animal species including cats, triggered the interest for the development of novel molecules with antiviral activity for treatment of CoV infections in humans and animals. Essential oils (EOs) have raised significant attention for their antiviral properties integrating and, in some cases, replacing conventional drugs. Thymus vulgaris EO (TEO) has been previously shown to be effective against several RNA viruses including CoVs. In the present study the antiviral efficacy of TEO against FIPV was evaluated in vitro. TEO at 27 µg/ml was able to inhibit virus replication with a significant reduction of 2 log10 TCID50/50 µl. Moreover, virucidal activity was tested using TEO at 27 and 270 µg/ml, over the cytotoxic threshold, determining a reduction of viral titre as high as 3.25 log10 TCID50/50 µl up to 1 h of time contact. These results open several perspectives in terms of future applications and therapeutic possibilities for coronaviruses considering that FIPV infection in cats could be a potential model for the study of antivirals against CoVs.