Armoured exogenous internal control for real-time PCR diagnosis of avian influenza

被引：4

作者：

Andreychuk, D. B. ^{[1
]}

Andriyasov, A., V ^{[1
]}

Nikonova, Z. B. ^{[1
]}

Kozlov, A. A. ^{[1
]}

Suarez, D. L. ^{[2
]}

Chvala, Il A. ^{[1
]}

机构：

[1] Fed Ctr Anim Hlth, Vladimir, Russia

[2] Us Natl Poultry Res Ctr, Southeast Poultry Res Lab, Athens, GA USA

来源：

AVIAN PATHOLOGY | 2019年 / 48卷 / 05期

关键词：

Real-time PCR; exogenous internal armoured PCR control; EGFP; avian influenza; PCR inhibitors; REVERSE TRANSCRIPTION-PCR; RNA; VIRUS; ASSAY; CONSTRUCTION; DELIVERY; GENE;

D O I：

10.1080/03079457.2019.1628918

中图分类号：

S85 [动物医学（兽医学）];

学科分类号：

0906 ;

摘要：

An exogenous "armoured" 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 increased physical stability of the IC was ensured using branched polyethyleneimine (PEI) which was in complex with IC-RNA. The option to add the IC directly to pathological material suspensions allows measurement of the nucleic acids extraction efficiency. Stability of armoured RNA-IC during storage and tissue suspension preparation was shown. The advantage of exogenous "armoured" 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 values in all tests.

引用

收藏

页码：492 / 498

页数：7

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