Exotic and Emerging Avian Viral Diseases Unit, US National Poultry Research Center, US Department of Agriculture, Agricultural Research Service, Athens, GA, USA.
Methods Mol Biol. 2020;2123:123-135. doi: 10.1007/978-1-0716-0346-8_10.
The efficient extraction and purification of viral RNA is critical for downstream molecular applications such as the sensitive and specific detection of virus in clinical samples, virus gene cloning and expression, gene sequencing, or quantification of avian influenza (AI) virus by molecular methods from experimentally infected birds. Samples can generally be divided into two types: enriched (e.g., virus stocks) and non-enriched (e.g., clinical). Clinical samples, which may be tissues or swab material, are the most difficult to process due to the complex sample composition and possibly low virus titers. In this chapter, two well-established procedures for the extraction of AI virus RNA from common clinical specimen types and enriched virus stocks will be presented.
从实验感染的鸟类中,通过分子方法对临床样本中的病毒进行敏感和特异性检测、病毒基因克隆和表达、基因测序或禽流感 (AI) 病毒的定量,病毒 RNA 的高效提取和纯化对于下游分子应用至关重要。样本通常可以分为两种类型:富集的(例如病毒株)和非富集的(例如临床样本)。由于样本组成复杂且病毒滴度可能较低,因此临床样本(可能是组织或拭子材料)是最难处理的。本章将介绍两种从常见临床标本类型和富集病毒株中提取 AI 病毒 RNA 的成熟程序。