Vaccine selection for rRT-PCR

Vaccine sample that showed the longest period of thermostability at 37°C was used for real-time PCR experiment. The PCR was based on a 150 bp, conserved region of the APMV-1 L polymerase gene, in which two probes were included, as per the protocol followed at the UK Animal and Plant Health Agency. Two serial dilutions of the vaccine were made: 10-fold and 2-fold. 10-fold serial dilution was made up to 10⁻³ and 2-fold up to 2⁻⁴. From each of 10-fold diluted sample, the sample was tested once for detecting the cycle threshold (C_T) value, while each 2-fold dilution was tested twice to calculate the mean C_T value.