RT-nested PCR for detection of HEV

The detection of HEV by RT-nested PCR was performed according to the protocol previously proposed [16] using primers for the region ORF1 of HEV. The reaction had a final volume of 50 μL containing 25 μL of Promega GoTaq Green Master Mix, 18 μL of water DNase/RNase-free, 1 ul of each primer in the contraction of 20 pmol, and 5 μL of cDNA. For amplification, the program was used with initial temperature of 95 °C for 5 min, followed by 45 cycles of 95 °C for 30 s, 59 °C for 1 min, 72 °C for 1 min, and at the end of the cycles 72 °C for 7 min. The second running was performed under the same conditions. After, the reactions of the electrophoresis of the amplified products were done, and the results were visualized with UV light.