The Quantitect One - Step RT - PCR kit (Qiagen, Hilden, Germany) was used with a 25 μl reaction
mixture under the following conditions: 0.25 μl of kit enzyme mixture (including reverse transcriptase RT and Taq polymerase), 10 μl of 2 × Quantitect RT - PCR buffer, 1.25 μl of 10 μM of each primer, 0.5 μl of 10 μM of probe at 10 μM, 6.8 μl of DNA RNA free water (Mol Bio grade, Hamburg, Germany) and 5 μl of the extracted sample.