Novel Corona virus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or 2019-nCoV), and the subsequent disease caused by the virus (coronavirus disease 2019 or COVID-19), is an emerging global health concern that requires a rapid diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the standard for SARS-CoV-2 detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper field based testing at point-of-risk. The objective of this study was to develop a rapid screening diagnostic test that could be completed in 30-45 minutes. Simulated patient samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the SARS-CoV-2 nucleic sequence. RNA isolated from nasopharyngeal swabs collected from actual COVID-19 patients was also tested. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses. RT-LAMP specifically detected SARS-CoV-2 in both simulated patient samples and clinical specimens. This test was performed in 30-45 minutes. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts in the field and potential ports of entry.

In vivo in the prostate gland, basal epithelial cells adhere to laminin 5 (LM5) via ␣3␤1 and ␣6␤4 integrins. When placed in culture primary prostate basal epithelial cells secrete and adhere to their own LM5-rich matrix. Adhesion to LM5 is required for cell survival that is dependent on integrin-mediated, ligand-independent activation of the epidermal growth factor receptor (EGFR) and the cytoplasmic tyrosine kinase Src, but not PI-3K. Integrin-mediated adhesion via ␣3␤1, but not ␣6␤4 integrin, supports cell survival through EGFR by signaling downstream to Erk. PC3 cells, which do not activate EGFR or Erk on LM5-rich matrices, are not dependent on this pathway for survival. PC3 cells are dependent on PI-3K for survival and undergo caspase-dependent death when PI-3K is inhibited. The death induced by inhibition of EGFR or Src in normal primary prostate cells is not mediated through or dependent on caspase activation, but depends on the induction of reactive oxygen species. In addition the presence of an autophagic pathway, maintained by adhesion to matrix through ␣3␤1 and ␣6␤4, prevents the induction of caspases when EGFR or Src is inhibited. Suppression of autophagy is sufficient to induce caspase activation and apoptosis in LM5-adherent primary prostate epithelial cells. INTRODUCTIONIn vivo, the precise regulation of epithelial cell homeostasis involves interactions between cells and their microenvironment. Cells receive signals from both the extracellular matrix in the basement membrane and soluble factors secreted by the stroma that precisely control the timing of cell division, growth arrest, differentiation, and survival. Integrins on the cell surface that interact with laminin 5 (LM5) in the extracellular matrix, such as ␣3␤1 and ␣6␤4, are critically involved in mediating survival. Genetic loss of LM5, or its receptors ␣3, ␣6, or ␤4 integrins, in vivo results in cell detachment and induction of caspase-mediated apoptosis, even in the presence of soluble factors (Ryan et al., 1999;DiPersio et al., 2000). This detachment-induced form of apoptosis has been termed anoikis (Frisch and Screaton, 2001). In vitro anoikis can be rescued by expression of an activated form of FAK, Rac, or Akt (Frisch et al., 1996;Rytomaa et al., 2000;Coniglio et al., 2001), suggesting that integrin-mediated signaling through these molecules is required to maintain cell survival. However, studies in which specific signaling pathways are inhibited while integrins are still engaged suggest alternative pathways, such as Ras/Erk or Jnk, are required for integrin-mediated survival (Almeida et al., 2000;Manohar et al., 2004). Whether signaling from multiple pathways is involved in mediating integrin-dependent survival and whether different pathways are unique to specific cell types have not been extensively investigated.In addition to classical caspase-mediated apoptosis, such as that observed during anoikis, several other mechanisms of cell death have been described (Melino et al., 2005). Other forms of cell death include caspase-independe...

Novel Corona virus (COVID-19 or 2019-nCoV) is an emerging global health concern that requires a rapid diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the standard for COVID-19 detection; however, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for faster and cheaper field based testing at point-of-risk. The objective of this study was to develop a rapid screening diagnostic test that could be completed in under 30 minutes. Simulated patient samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the COVID-19 nucleic sequence. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses. RT-LAMP specifically detected COVID-19 in simulated patient samples.This test was performed in under 30 minutes. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts in the field and potential ports of entry.All rights reserved. No reuse allowed without permission. author/funder, who has granted medRxiv a license to display the preprint in perpetuity.