Armored Exogenous Internal Control for Real-Time PCR Diagnosis of Avian Influenza

An exogenous "armored" PCR internal control (IC) short RNA was analyzed in conjunction with real-time RT-PCR method for diagnosis of avian influenza. The resistance to nucleases and increase the physical stability of the IC was ensured branched polyethyleneimine (PEI) which was in complex with IC-RNA. The option to add the IC directly to pathological material suspensions allows to measure the nucleic acids extraction efficiency. Stability of armored RNA-IC during storage and tissue suspension preparation was shown. The advantage of exogenous "armored" IC was demonstrated in the experiment with AIV genome detection by QPCR in samples from different species of wild birds. The exogenous IC gave reproducible homogeneous Ct value in all tests.