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Avian Pathol. 2019 Jun 17:1-15. doi: 10.1080/03079457.2019.1628918. [Epub ahead of print]
Armored Exogenous Internal Control for Real-Time PCR Diagnosis of Avian Influenza.
Andreychuk DB1, Andriyasov AV1, Nikonova ZB1, Kozlov AA1, Suarez DL2, Chvala IA1.
Author information
Abstract
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.
KEYWORDS:
EGFP; PCR inhibitors; avian influenza; exogenous internal armored PCR control; real-time PCR
PMID: 31203638 DOI: 10.1080/03079457.2019.1628918
Armored Exogenous Internal Control for Real-Time PCR Diagnosis of Avian Influenza.
Andreychuk DB1, Andriyasov AV1, Nikonova ZB1, Kozlov AA1, Suarez DL2, Chvala IA1.
Author information
Abstract
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.
KEYWORDS:
EGFP; PCR inhibitors; avian influenza; exogenous internal armored PCR control; real-time PCR
PMID: 31203638 DOI: 10.1080/03079457.2019.1628918