Internal reference genes with the potential for normalizing quantitative PCR results for oral fluid specimens.

In basic research, testing of oral fluid specimens by real-time quantitative polymerase chain reaction (qPCR) has been used to evaluate changes in gene expression levels following experimental treatments. In diagnostic medicine, qPCR has been used to detect DNA/RNA transcripts indicative of bacterial or viral infections. Normalization of qPCR using endogenous and exogenous reference genes is a well-established strategy for ensuring result comparability by controlling sample-to-sample variation introduced during sampling, storage, and qPCR testing. In this review, the majority of recent publications in human (n = 136) and veterinary (n = 179) medicine did not describe the use of internal reference genes in qPCRs for oral fluid specimens (52.9% animal studies; 57.0% human studies). However, the use of endogenous reference genes has not been fully explored or validated for oral fluid specimens. The lack of valid internal reference genes inherent to the oral fluid matrix will continue to hamper the reliability, reproducibility, and generalizability of oral fluid qPCR assays until this issue is addressed.

Evaluation of an Air Cleaning Device Equipped with Filtration and UV: Comparison of Removal Efficiency on Particulate Matter and Viable Airborne Bacteria in the Inlet and Treated Air.

Since the COVID-19 pandemic, improving indoor air quality (IAQ) has become vital for the public as COVID-19 and other infectious diseases can transmit via inhalable aerosols. Air cleaning devices with filtration and targeted pollutant treatment capabilities can help improve IAQ. However, only a few filtration/UV devices have been formally tested for their effectiveness, and little data is publicly available and UV doses comparable. In this research, we upgraded a particulate matter (PM) air filtration prototype by adding UV-C (germicidal) light. We developed realistic UV dose metrics for fast-moving air and selected performance scenarios to quantify the mitigation effect on viable airborne bacteria and PM. The targeted PM included total suspended particulate (TSP) and a coarse-to-fine range sized at PM10, PM4, PM2.5, and PM1. The PM and viable airborne bacteria concentrations were compared between the inlet and outlet of the prototype at 0.5 and 1.0 m3/s (low and high) air flow modes. The upgraded prototype inactivated nearly 100% of viable airborne bacteria and removed up to 97% of TSP, 91% of PM10, 87% of PM4, 87% of PM2.5, and 88% of PM1. The performance in the low flow rate mode was generally better than in the high flow rate mode. The combination of filtration and UV-C treatment provided 'double-barrier' assurance for air purification and lowered the risk of spreading infectious micro-organisms.

The N-terminal Subunit of the Porcine Deltacoronavirus Spike Recombinant Protein (S1) Does Not Serologically Cross-react with Other Porcine Coronaviruses.

Porcine deltacoronavirus (PDCoV), belonging to family Coronaviridae and genus Deltacoronavirus, is a major enteric pathogen in swine. Accurate PDCoV diagnosis relying on laboratory testing and antibody detection is an important approach. This study evaluated the potential of the receptor-binding subunit of the PDCoV spike protein (S1), generated using a mammalian expression system, for specific antibody detection via indirect enzyme-linked immunosorbent assay (ELISA). Serum samples were collected at day post-inoculation (DPI) -7 to 42, from pigs (n = 83) experimentally inoculated with different porcine coronaviruses (PorCoV). The diagnostic sensitivity of the PDCoV S1-based ELISA was evaluated using serum samples (n = 72) from PDCoV-inoculated animals. The diagnostic specificity and potential cross-reactivity of the assay was evaluated on PorCoV-negative samples (n = 345) and samples collected from pigs experimentally inoculated with other PorCoVs (n = 472). The overall diagnostic performance, time of detection, and detection rate over time varied across different S/P cut-offs, estimated by Receiver Operating Characteristic (ROC) curve analysis. The higher detection rate in the PDCoV group was observed after DPI 21. An S/P cut-off of 0.25 provided 100% specificity with no serological cross-reactivity against other PorCoV. These results support the use of S1 protein-based ELISA for accurate detection of PDCoV infections, transference of maternal antibodies, or active surveillance.