Spackman Erica, Suarez David L
Southeast Poultry Research Laboratory, US Department of Agriculture, Agricultural Research Service, Athens, GA, USA.
Methods Mol Biol. 2008;436:13-8. doi: 10.1007/978-1-59745-279-3_3.
The efficient extraction and purification of viral RNA are critical for downstream molecular applications, whether it is the sensitive and specific detection of virus in clinical samples, virus gene cloning and expression, or quantification of the 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 clinical. Clinical type 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 isolation of AI virus RNA from common clinical specimen types and enriched virus stocks for further molecular applications will be presented.
病毒RNA的高效提取和纯化对于下游分子应用至关重要,无论是临床样本中病毒的灵敏特异检测、病毒基因克隆与表达,还是通过分子方法对实验感染禽类的禽流感(AI)病毒进行定量分析。样本通常可分为两类:富集样本(如病毒储备液)和临床样本。临床类样本可能是组织或拭子材料,由于样本组成复杂且病毒滴度可能较低,是最难处理的。在本章中,将介绍两种成熟的从常见临床样本类型和富集病毒储备液中分离AI病毒RNA以用于进一步分子应用的方法。