Fig. 2

A Optimization of the triplex primer concentration for amplification. PCR amplification was performed using different concentrations of a triplex primer cocktail composed of forward and reverse primer sets for MERS-CoV, NiV and REBOV in equimolar ratios. Lane a - 1.2 µM, Lane b - 0.8 µM, Lane c - 0.6 µM, Lane d - 0.4 µM, Lane e - 0.3 µM, Lane f - 200 nM, Lane g - 150 nM, Lane h - 100 nM. Ladder - 50 bp. B Optimization of the annealing temperature for amplification. PCR optimization using different annealing temperatures for triplex amplification with the optimized triplex primer cocktail. Lane a - MERS-CoV (92 bp), lane b - NiV (100 bp), lane c - REBOV (80 bp), and lane d - NTC. Ladder - 50 bp