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The Shannon-Weaver diversity index (H) and richness index (S) were calculated using an OD of 0.25 as threshold for positive response, which was described by Garland and Derry et al. [ 29, 30]: (2) H = − ∑ p i ln p i, where p i is the ratio of the activity on each substrate to the sum of activities on all substrates.
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When we used an OD cutoff of 1.0, these values were 9.0%, 44.9%, and 4.7% respectively.
Using a cutoff OD of 0.19, all 129 NPAs positive for respiratory viruses showed very low OD, with a specificity of 100%.
Cell density was measured using a spectrophotometer at an OD of 600 nm.
Briefly, a 10 ml culture in Mueller-Hinton (MH) broth containing 0.1% glucose, 0.025% ferric pyrophosphate, and 2% IsoVitaleX (Becton Dickinson) was inoculated to an OD of ∼0.15 using overnight cultures of each of the deletion mutants.
The antibody titer was calculated by extrapolating the serum dilution corresponding to an OD of 0.2 using linear regression.
An amount of 1.5-10 1.5-10 determined using an electrophotometer with an OD 260/280 ratio of at least 1.8 and gel electrophoresis.
Positive responses were defined as an OD of 2x the mean OD of healthy endemic controls.
The optical density values were normalized in every run using a calibrator (normalized OD (ODn) = mean specimen OD/mean calibrator OD).
The SOD was calculated using the median OD of the triplicates.
The proliferation curve was generated using the equation: OD of hybrids/OD of control cells × 100%.
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