Quantitative fluorescence experiments were performed in parallel with reactions incubated in both the test device and in the ESE-Quant Tube Scanner, programmed for a constant incubation at 63uC. For NINA incubated reactions, LAMP was terminated at the time (,36 min.) when the signal from the parallel reactions on the Tube Scanner began to indicate detectable amplification to avoid signal saturation in all dilutions. Termination was accomplished by flash chilling and later inactivating the reaction by heating at 80uC for 5 minutes. The fluorescence signals of the NINA incubated samples were then read on the SpectraMax M2 plate reader with l ex = 485 nm and l em = 515 nm. Figure S1 The workflow of a proposed NA amplification assay kit. The kit will be an instrument-free, electricity-free nucleic-acid amplification test that is compatible with whole blood, is temperature stable and contains contamination. 1) Initiate NINA heater by installing heater cartridge (a) into insulated housing (b), add EPCM module (c) and lid (d). 2) Set up for assay by opening single assay subkit. 3) Sample blood to calibrated line on collection capillary. 4) Transfer blood and blister contents to ''S'' tube and prefilled diluent to ''NC'' and ''PC'' tubes and mix all. 5) Amplify. Verify temperature ''ready'' indication on the NINA device through transparent view port in the lid, remove the lid, add the three tubes to the NINA heater, and replace lid. Incubate 45 minutes. Verify temperature is still in range through transparent view port (process control). 6) Quench to all three tubes by pushing cap to burst frangible seal and transfer ,10 mL diluted quencher to the amplified mixture. (TIF)  Quantitative Subcellular Proteome and Secretome Profiling of Influenza A Virus-Infected Human Primary Macrophages Influenza A viruses are important pathogens that cause acute respiratory diseases and annual epidemics in humans. Macrophages recognize influenza A virus infection with their pattern recognition receptors, and are involved in the activation of proper innate immune response. Here, we have used high-throughput subcellular proteomics combined with bioinformatics to provide a global view of host cellular events that are activated in response to influenza A virus infection in human primary macrophages. We show that viral infection regulates the expression and/or subcellular localization of more than one thousand host proteins at early phases of infection. Our data reveals that there are dramatic changes in mitochondrial and nuclear proteomes in response to infection. We show that a rapid cytoplasmic leakage of lysosomal proteins, including cathepsins, followed by their secretion, contributes to inflammasome activation and apoptosis seen in the infected macrophages. Also, our results demonstrate that P2X(7) receptor and src tyrosine kinase activity are essential for inflammasome activation during influenza A virus infection. Finally, we show that influenza A virus infection is associated with robust secretion of different danger-associated molecular patterns (DAMPs) suggesting an important role for DAMPs in host response to influenza A virus infection. In conclusion, our high-throughput quantitative proteomics study provides important new insight into host-response against influenza A virus infection in human primary macrophages. Influenza A viruses are negative-stranded RNA viruses that are capable of infecting a variety of avian and mammalian species. These viruses are responsible for the annual epidemics that cause severe illnesses in millions of people worldwide. Influenza A virus and secondary bacterial infections can cause lethal pneumonia and encephalopathy especially in elder people. Host defence against influenza A virus infection is initiated by the innate immune system. The principal effector cells involved in innate immunity are macrophages, and dendritic cells (DC) that kill microbes through phagocytosis, present antigens to T cells, and produce cytokines. These innate immune responses are essential for the development of later adaptive immune responses, which provide specific cell-mediated and humoral protection, and are often necessary for a complete clearance of infection.